本(ben)(ben)試(shi)(shi)劑盒(he)(he)(he)只能用(yong)于(yu)科學(xue)研(yan)究(jiu)，不(bu)(bu)得(de)用(yong)于(yu)醫學(xue)診斷(duan)人（Human）轉(zhuan)(zhuan)(zhuan)化(hua)(hua)(hua)生長因子β2（TGF-β2）ELISA檢(jian)測(ce)(ce)(ce)試(shi)(shi)劑盒(he)(he)(he)使(shi)(shi)(shi)用(yong)說(shuo)(shuo)(shuo)明(ming)(ming)書(shu)檢(jian)測(ce)(ce)(ce)原(yuan)試(shi)(shi)劑盒(he)(he)(he)采用(yong)雙抗(kang)體(ti)(ti)一(yi)步夾心(xin)法酶(mei)(mei)聯免疫吸附試(shi)(shi)驗(yan)（ELISA）。往預(yu)(yu)先(xian)包(bao)被轉(zhuan)(zhuan)(zhuan)化(hua)(hua)(hua)生長因子β2（TGF-β2）抗(kang)體(ti)(ti)的(de)(de)(de)包(bao)被微孔(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)(zhong)(zhong)(zhong)，依次(ci)加(jia)(jia)入(ru)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)本(ben)(ben)、標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)、HRP標(biao)(biao)(biao)(biao)(biao)(biao)(biao)記(ji)(ji)的(de)(de)(de)檢(jian)測(ce)(ce)(ce)抗(kang)體(ti)(ti)，經過(guo)溫(wen)(wen)(wen)育(yu)并徹底(di)洗(xi)(xi)(xi)(xi)滌(di)。用(yong)底(di)物(wu)TMB顯色，TMB在(zai)過(guo)氧化(hua)(hua)(hua)物(wu)酶(mei)(mei)的(de)(de)(de)催化(hua)(hua)(hua)下(xia)轉(zhuan)(zhuan)(zhuan)化(hua)(hua)(hua)成藍色，并在(zai)酸的(de)(de)(de)作(zuo)(zuo)用(yong)下(xia)轉(zhuan)(zhuan)(zhuan)化(hua)(hua)(hua)成＊終的(de)(de)(de)黃色。顏色的(de)(de)(de)深淺和(he)(he)樣(yang)(yang)(yang)(yang)品(pin)(pin)中(zhong)(zhong)(zhong)(zhong)(zhong)的(de)(de)(de)轉(zhuan)(zhuan)(zhuan)化(hua)(hua)(hua)生長因子β2（TGF-β2）呈正(zheng)相(xiang)(xiang)關(guan)。用(yong)酶(mei)(mei)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)儀在(zai)450nm 波(bo)長下(xia)測(ce)(ce)(ce)定吸光(guang)(guang)度(du)(du)（OD 值），計(ji)算(suan)樣(yang)(yang)(yang)(yang)品(pin)(pin)濃(nong)(nong)度(du)(du)。樣(yang)(yang)(yang)(yang)品(pin)(pin)收(shou)集、處(chu)(chu)理及(ji)保存方(fang)(fang)(fang)法1. 血(xue)清：使(shi)(shi)(shi)用(yong)不(bu)(bu)含熱原(yuan)和(he)(he)內(nei)(nei)毒素的(de)(de)(de)試(shi)(shi)管(guan)，操(cao)作(zuo)(zuo)過(guo)程(cheng)中(zhong)(zhong)(zhong)(zhong)(zhong)避(bi)免任何(he)細(xi)胞刺(ci)激，收(shou)集血(xue)液(ye)后(hou)，3000轉(zhuan)(zhuan)(zhuan)離(li)(li)(li)心(xin)10分(fen)(fen)(fen)鐘將血(xue)清和(he)(he)紅細(xi)胞迅速小(xiao)心(xin)地分(fen)(fen)(fen)離(li)(li)(li)。2. 血(xue)漿：EDTA、檸檬酸鹽或肝(gan)素抗(kang)凝(ning)。3000轉(zhuan)(zhuan)(zhuan)離(li)(li)(li)心(xin)30分(fen)(fen)(fen)鐘取(qu)上清。3. 細(xi)胞上清液(ye)：3000轉(zhuan)(zhuan)(zhuan)離(li)(li)(li)心(xin)10分(fen)(fen)(fen)鐘去(qu)除(chu)顆粒和(he)(he)聚(ju)合物(wu)。4. 組(zu)(zu)(zu)織勻漿：將組(zu)(zu)(zu)織加(jia)(jia)入(ru)適量(liang)(liang)生理鹽水(shui)(shui)(shui)搗碎(sui)。3000轉(zhuan)(zhuan)(zhuan)離(li)(li)(li)心(xin)10分(fen)(fen)(fen)鐘取(qu)上清。5. 保存：如(ru)果(guo)(guo)樣(yang)(yang)(yang)(yang)本(ben)(ben)收(shou)集后(hou)不(bu)(bu)及(ji)時檢(jian)測(ce)(ce)(ce)，請按(an)(an)(an)一(yi)次(ci)用(yong)量(liang)(liang)分(fen)(fen)(fen)裝，凍(dong)存于(yu)-20℃，避(bi)免反復(fu)凍(dong)融，在(zai)室(shi)溫(wen)(wen)(wen)下(xia)解凍(dong)并確保樣(yang)(yang)(yang)(yang)品(pin)(pin)均勻地充分(fen)(fen)(fen)解凍(dong)。自備物(wu)品(pin)(pin)1. 酶(mei)(mei)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)儀（450nm）2. 高(gao)精度(du)(du)加(jia)(jia)樣(yang)(yang)(yang)(yang)器及(ji)槍頭：0.5-10uL、2-20uL、20-200uL、200-1000uL3. 37℃恒(heng)溫(wen)(wen)(wen)箱操(cao)作(zuo)(zuo)注意(yi)事項1. 試(shi)(shi)劑盒(he)(he)(he)保存在(zai)2-8℃，使(shi)(shi)(shi)用(yong)前室(shi)溫(wen)(wen)(wen)平衡20分(fen)(fen)(fen)鐘。從冰箱取(qu)出的(de)(de)(de)濃(nong)(nong)縮洗(xi)(xi)(xi)(xi)滌(di)液(ye)會有結(jie)晶，這屬于(yu)正(zheng)常(chang)現(xian)象，水(shui)(shui)(shui)浴(yu)加(jia)(jia)熱使(shi)(shi)(shi)結(jie)晶完全(quan)溶(rong)解后(hou)再使(shi)(shi)(shi)用(yong)。2. 實驗(yan)中(zhong)(zhong)(zhong)(zhong)(zhong)不(bu)(bu)用(yong)的(de)(de)(de)板(ban)(ban)(ban)(ban)(ban)條(tiao)應立(li)即(ji)放(fang)回(hui)(hui)自封(feng)(feng)袋(dai)中(zhong)(zhong)(zhong)(zhong)(zhong)，密(mi)封(feng)(feng)（低溫(wen)(wen)(wen)干燥）保存。3. 濃(nong)(nong)度(du)(du)為(wei)(wei)0的(de)(de)(de)S0號標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)即(ji)可視(shi)為(wei)(wei)陰(yin)性(xing)(xing)(xing)對照或者(zhe)(zhe)空(kong)白(bai)；按(an)(an)(an)照說(shuo)(shuo)(shuo)明(ming)(ming)書(shu)操(cao)作(zuo)(zuo)時樣(yang)(yang)(yang)(yang)本(ben)(ben)已經稀釋(shi)5倍，＊終結(jie)果(guo)(guo)乘以(yi)5才是樣(yang)(yang)(yang)(yang)本(ben)(ben)實際濃(nong)(nong)度(du)(du)。4. 嚴格按(an)(an)(an)照說(shuo)(shuo)(shuo)明(ming)(ming)書(shu)中(zhong)(zhong)(zhong)(zhong)(zhong)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)明(ming)(ming)的(de)(de)(de)時間(jian)、加(jia)(jia)液(ye)量(liang)(liang)及(ji)順序進行溫(wen)(wen)(wen)育(yu)操(cao)作(zuo)(zuo)。5. 所有液(ye)體(ti)(ti)組(zu)(zu)(zu)分(fen)(fen)(fen)使(shi)(shi)(shi)用(yong)前充分(fen)(fen)(fen)搖(yao)勻。試(shi)(shi)劑盒(he)(he)(he)組(zu)(zu)(zu)成名(ming)稱96孔(kong)(kong)(kong)(kong)(kong)配置(zhi)48孔(kong)(kong)(kong)(kong)(kong)配置(zhi)備注微孔(kong)(kong)(kong)(kong)(kong)酶(mei)(mei)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)板(ban)(ban)(ban)(ban)(ban)12孔(kong)(kong)(kong)(kong)(kong)×8條(tiao)12孔(kong)(kong)(kong)(kong)(kong)×4條(tiao)無(wu)(wu)(wu)(wu)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)0.3mL*6管(guan)0.3mL*6管(guan)無(wu)(wu)(wu)(wu)樣(yang)(yang)(yang)(yang)本(ben)(ben)稀釋(shi)液(ye)6mL3mL無(wu)(wu)(wu)(wu)檢(jian)測(ce)(ce)(ce)抗(kang)體(ti)(ti)-HRP10mL5mL無(wu)(wu)(wu)(wu)20×洗(xi)(xi)(xi)(xi)滌(di)緩(huan)沖液(ye)25mL15mL按(an)(an)(an)說(shuo)(shuo)(shuo)明(ming)(ming)書(shu)進行稀釋(shi)底(di)物(wu)A6mL3mL無(wu)(wu)(wu)(wu)底(di)物(wu)B6mL3mL無(wu)(wu)(wu)(wu)終止液(ye)6mL3mL無(wu)(wu)(wu)(wu)封(feng)(feng)板(ban)(ban)(ban)(ban)(ban)膜2張(zhang)(zhang)2張(zhang)(zhang)無(wu)(wu)(wu)(wu)說(shuo)(shuo)(shuo)明(ming)(ming)書(shu)1份(fen)(fen)(fen)1份(fen)(fen)(fen)無(wu)(wu)(wu)(wu)自封(feng)(feng)袋(dai)1個1個無(wu)(wu)(wu)(wu)注：標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)（S0-S5）濃(nong)(nong)度(du)(du)依次(ci)為(wei)(wei)：0、250、500、1000、2000、4000 pg/mL試(shi)(shi)劑的(de)(de)(de)準(zhun)(zhun)(zhun)(zhun)備 20×洗(xi)(xi)(xi)(xi)滌(di)緩(huan)沖液(ye)的(de)(de)(de)稀釋(shi)：蒸(zheng)餾水(shui)(shui)(shui)按(an)(an)(an)1：20稀釋(shi)，即(ji)1份(fen)(fen)(fen)的(de)(de)(de)20×洗(xi)(xi)(xi)(xi)滌(di)緩(huan)沖液(ye)加(jia)(jia)19份(fen)(fen)(fen)的(de)(de)(de)蒸(zheng)餾水(shui)(shui)(shui)。洗(xi)(xi)(xi)(xi)板(ban)(ban)(ban)(ban)(ban)方(fang)(fang)(fang)法1. 手工洗(xi)(xi)(xi)(xi)板(ban)(ban)(ban)(ban)(ban)：甩(shuai)盡孔(kong)(kong)(kong)(kong)(kong)內(nei)(nei)液(ye)體(ti)(ti)，每(mei)孔(kong)(kong)(kong)(kong)(kong)加(jia)(jia)滿洗(xi)(xi)(xi)(xi)滌(di)液(ye)，靜置(zhi)1min后(hou)甩(shuai)盡孔(kong)(kong)(kong)(kong)(kong)內(nei)(nei)液(ye)體(ti)(ti)，在(zai)吸水(shui)(shui)(shui)紙(zhi)上拍(pai)干，如(ru)此洗(xi)(xi)(xi)(xi)板(ban)(ban)(ban)(ban)(ban)5次(ci)。2. 自動洗(xi)(xi)(xi)(xi)板(ban)(ban)(ban)(ban)(ban)機(ji)：每(mei)孔(kong)(kong)(kong)(kong)(kong)注入(ru)洗(xi)(xi)(xi)(xi)液(ye)350μL，浸泡1min，洗(xi)(xi)(xi)(xi)板(ban)(ban)(ban)(ban)(ban)5次(ci)。操(cao)作(zuo)(zuo)步驟(zou)1. 從室(shi)溫(wen)(wen)(wen)平衡20min后(hou)的(de)(de)(de)鋁箔袋(dai)中(zhong)(zhong)(zhong)(zhong)(zhong)取(qu)出所需板(ban)(ban)(ban)(ban)(ban)條(tiao)，剩余板(ban)(ban)(ban)(ban)(ban)條(tiao)用(yong)自封(feng)(feng)袋(dai)密(mi)封(feng)(feng)放(fang)回(hui)(hui)4℃。2. 設(she)置(zhi)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)孔(kong)(kong)(kong)(kong)(kong)和(he)(he)樣(yang)(yang)(yang)(yang)本(ben)(ben)孔(kong)(kong)(kong)(kong)(kong)，標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)孔(kong)(kong)(kong)(kong)(kong)各(ge)加(jia)(jia)不(bu)(bu)同(tong)濃(nong)(nong)度(du)(du)的(de)(de)(de)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)50μL；3. 樣(yang)(yang)(yang)(yang)本(ben)(ben)孔(kong)(kong)(kong)(kong)(kong)先(xian)加(jia)(jia)待測(ce)(ce)(ce)樣(yang)(yang)(yang)(yang)本(ben)(ben)10μL，再加(jia)(jia)樣(yang)(yang)(yang)(yang)本(ben)(ben)稀釋(shi)液(ye)40μL；空(kong)白(bai)孔(kong)(kong)(kong)(kong)(kong)不(bu)(bu)加(jia)(jia)。 4. 除(chu)空(kong)白(bai)孔(kong)(kong)(kong)(kong)(kong)外，標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)孔(kong)(kong)(kong)(kong)(kong)和(he)(he)樣(yang)(yang)(yang)(yang)本(ben)(ben)孔(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)(zhong)(zhong)(zhong)每(mei)孔(kong)(kong)(kong)(kong)(kong)加(jia)(jia)入(ru)辣根過(guo)氧化(hua)(hua)(hua)物(wu)酶(mei)(mei)（HRP）標(biao)(biao)(biao)(biao)(biao)(biao)(biao)記(ji)(ji)的(de)(de)(de)檢(jian)測(ce)(ce)(ce)抗(kang)體(ti)(ti)100μL，用(yong)封(feng)(feng)板(ban)(ban)(ban)(ban)(ban)膜封(feng)(feng)住反應孔(kong)(kong)(kong)(kong)(kong)，37℃水(shui)(shui)(shui)浴(yu)鍋或恒(heng)溫(wen)(wen)(wen)箱溫(wen)(wen)(wen)育(yu)60min。5. 棄去(qu)液(ye)體(ti)(ti)，吸水(shui)(shui)(shui)紙(zhi)上拍(pai)干，每(mei)孔(kong)(kong)(kong)(kong)(kong)加(jia)(jia)滿洗(xi)(xi)(xi)(xi)滌(di)液(ye)，靜置(zhi)1min，甩(shuai)去(qu)洗(xi)(xi)(xi)(xi)滌(di)液(ye)，吸水(shui)(shui)(shui)紙(zhi)上拍(pai)干，如(ru)此重復(fu)洗(xi)(xi)(xi)(xi)板(ban)(ban)(ban)(ban)(ban)5次(ci)（也可用(yong)洗(xi)(xi)(xi)(xi)板(ban)(ban)(ban)(ban)(ban)機(ji)洗(xi)(xi)(xi)(xi)板(ban)(ban)(ban)(ban)(ban)）。6. 每(mei)孔(kong)(kong)(kong)(kong)(kong)加(jia)(jia)入(ru)底(di)物(wu)A、B各(ge)50μL，37℃避(bi)光(guang)(guang)孵育(yu)15min。7. 每(mei)孔(kong)(kong)(kong)(kong)(kong)加(jia)(jia)入(ru)終止液(ye)50μL，15min內(nei)(nei)，在(zai)450nm波(bo)長處(chu)(chu)測(ce)(ce)(ce)定各(ge)孔(kong)(kong)(kong)(kong)(kong)的(de)(de)(de)OD值。8.結(jie)果(guo)(guo)判斷(duan)9. 繪(hui)制標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)曲(qu)線：在(zai)Excel工作(zuo)(zuo)表中(zhong)(zhong)(zhong)(zhong)(zhong)，以(yi)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)濃(nong)(nong)度(du)(du)作(zuo)(zuo)橫坐(zuo)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)，對應OD值作(zuo)(zuo)縱坐(zuo)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)，繪(hui)制出標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)線性(xing)(xing)(xing)回(hui)(hui)歸(gui)曲(qu)線，按(an)(an)(an)曲(qu)線方(fang)(fang)(fang)程(cheng)計(ji)算(suan)各(ge)樣(yang)(yang)(yang)(yang)本(ben)(ben)濃(nong)(nong)度(du)(du)值。 試(shi)(shi)劑盒(he)(he)(he)性(xing)(xing)(xing)能1. 準(zhun)(zhun)(zhun)(zhun)確性(xing)(xing)(xing)：標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)線性(xing)(xing)(xing)回(hui)(hui)歸(gui)與預(yu)(yu)期濃(nong)(nong)度(du)(du)相(xiang)(xiang)關(guan)系(xi)(xi)數R值，大于(yu)等于(yu)0.9900。2. 靈敏度(du)(du)：＊低檢(jian)測(ce)(ce)(ce)濃(nong)(nong)度(du)(du)小(xiao)于(yu)50 pg/mL。3. 特異性(xing)(xing)(xing)：不(bu)(bu)與其它可溶(rong)性(xing)(xing)(xing)結(jie)構類似物(wu)交叉反應。4. 重復(fu)性(xing)(xing)(xing)：板(ban)(ban)(ban)(ban)(ban)內(nei)(nei)、板(ban)(ban)(ban)(ban)(ban)間(jian)變異系(xi)(xi)數均小(xiao)于(yu)15%。5. 貯(zhu)藏：2-8℃，避(bi)光(guang)(guang)防潮保存。6. 有效(xiao)期：6個月(yue)免責聲(sheng)明(ming)(ming)1. 試(shi)(shi)劑盒(he)(he)(he)僅供研(yan)究(jiu)使(shi)(shi)(shi)用(yong)，不(bu)(bu)得(de)用(yong)于(yu)臨床實驗(yan)或人體(ti)(ti)實驗(yan)，否(fou)則所產生的(de)(de)(de)一(yi)切(qie)后(hou)果(guo)(guo)，由實驗(yan)者(zhe)(zhe)承擔，本(ben)(ben)公司概不(bu)(bu)負責。2. 嚴格按(an)(an)(an)照說(shuo)(shuo)(shuo)明(ming)(ming)書(shu)操(cao)作(zuo)(zuo)，實驗(yan)者(zhe)(zhe)違反說(shuo)(shuo)(shuo)明(ming)(ming)書(shu)操(cao)作(zuo)(zuo)，后(hou)果(guo)(guo)由實驗(yan)者(zhe)(zhe)承擔。 FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES. Human Transforming Growth factor β2 (TGF-β2) ELISA Kit instruction Intended useThis TGF-β2 ELISA kit is intended Laboratory for Research use only and is not for use in diagnostic or therapeutic procedures.The Stop Solution changes the color from blue to yellow and the intensity of the color is measured at 450 nm using a spectrophotometer. In order to measure the concentration of TGF-β2 in the sample, this TGF-β2 ELISA Kit includes a set of calibration standards. The calibration standards are assayed at the same time as the samples and allow the operator to produce a standard curve of Optical Density versus TGF-β2 concentration. The concentration of TGF-β2 in the samples is then determined by comparing the O.D. of the samples to the standard curve.Sample collection and storagesSerum - Use a serum separator tube and allow samples to clot for 30 minutes before centrifugation for 10 minutes at approximately 3000×g. Remove serum and assay immediately or aliquot and store samples at -20℃ or -80℃.Avoid repeated freeze-thaw cyclesPlasma - Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 30 minutes at 3000×g at 2-8℃ within 30 minutes of collection. Store samples at -20℃or -80℃. Avoid repeated freeze-thaw cycles.Cell culture supernates and other biological fluids - Remove particulates by centrifugation and assay immediately or aliquot and store samples at -20℃or -80℃. Avoid repeated freeze-thaw cycles.Note: The samples shoule be centrifugated dequately and no hemolysis or granule was allowed.Materials required but not supplied1. Standard microplate reader(450nm)2. Precision pipettes and Disposable pipette tips.3. 37 ℃ incubatorPrecautions1. Do not substitute reagents from one kit to another. Standard, conjugate and microplates are matched for optimal performance. Use only the reagents supplied by manufacturer.2. Do not remove microplate from the storage bag until needed. Unused strips should be stored at 2-8°C in their pouch with the desiccant provided.3. Mix all reagents before using.Remove all kit reagents from refrigerator and allow them to reach room temperature ( 20-25°C)Materials suppliedName96 determinations48 determinationsMicroelisa stripplate12*8strips12*4stripsStandard0.3ml*6tubes0.3ml*6tubesSample Diluent6.0ml3.0mlHRP-Conjugate reagent10.0ml5.0ml20X Wash solution25ml15mlChromogen Solution A6.0ml3.0mlChromogen Solution B6.0ml3.0mlStop Solution6.0ml3.0mlClosure plate membrane22User manual11Sealed bags11Note: Standard (S0 → S5) concentration was followed by:0,250,500,1000,2000,4000 pg/mlReagent preparation20×wash solution:Dilute with Distilled or deionized water 1:20.Assay procedure1. Prepare all reagents before starting assay procedure. It is recommended that all Standards and Samples be added in duplicate to the Microelisa Stripplate.2. Add standard: Set Standard wells, testing sample wells. Add standard 50μl to standard well.3. Add Sample: Add testing sample 10μl then add Sample Diluent 40μl to testing sample well; Blank well doesn’t add anyting.4. Add 100μl of HRP-conjugate reagent to each well, cover with an adhesive strip and incubate for 60 minutes at 37°C. 5. Aspirate each well and wash, repeating the process four times for a total of five washes. Wash by filling each well with Wash Solution (400μl) using a squirt bottle, manifold dispenser or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Solution by aspirating or decanting. Invert the plate and blot it against clean paper towels.6. Add chromogen solution A 50μl and chromogen solution B 50μl to each well. Gently mix and incubate for 15 minutes at 37°C. Protect from light.7. Add 50μl Stop Solution to each well. The color in the wells should change from blue to yellow. If the color in the wells is green or the color change does not appear uniform, gently tap the plate to ensure thorough mixing.8. Read the Optical Density (O.D.) at 450 nm using a microtiter plate reader within 15 minutes.Calculation of results1. This standard curve is used to determine the amount in an unknown sample. The standard curve is generated by plotting the average O.D. (450 nm) obtained for each of the six standard concentrations on the vertical (Y) axis versus the corresponding concentration on the horizontal (X) axis. 2. First, calculate the mean O.D. value for each standard and sample. All O.D. values, are subtracted by the mean value of the zero standard before result interpretation. Construct the standard curve using graph paper or statistical software. 3. To determine the amount in each sample, first locate the O.D. value on the Y-axis and extend a horizontal line to the standard curve. At the point of intersection, draw a vertical line to the X-axis and read the corresponding concentration. 4. Any variation in operator, pipetting and washing technique, incubation time or temperature, and kit age can cause variation in result. Each user should obtain their own standard curve.5. The sensitivity by this assay is 50 pg/ml6. Standard curve Storage： 2-8℃.validity： six months. FOR RESEARCH USE ONLY; NOT FOR THERAPEUTIC OR DIAGNOSTIC APPLICATIONS! PLEASE READ THROUGH ENTIRE PROCEDURE BEFORE BEGINNING!
本(ben)(ben)(ben)(ben)試劑(ji)盒(he)(he)(he)(he)只能用(yong)(yong)(yong)(yong)(yong)(yong)于(yu)科學(xue)研(yan)究，不(bu)(bu)得(de)用(yong)(yong)(yong)(yong)(yong)(yong)于(yu)醫學(xue)診(zhen)斷(duan) 人(ren)(ren)（Human）尿(niao)膽素(su)(su)(su)(su)（urobilin）ELISA 檢(jian)(jian)測(ce)(ce)試劑(ji)盒(he)(he)(he)(he)使(shi)(shi)用(yong)(yong)(yong)(yong)(yong)(yong)說(shuo)(shuo)明(ming)書(shu) 檢(jian)(jian)測(ce)(ce)原理(li) 試劑(ji)盒(he)(he)(he)(he)采(cai)用(yong)(yong)(yong)(yong)(yong)(yong)雙抗(kang)(kang)體(ti)一步夾(jia)心(xin)(xin)法酶聯免(mian)疫吸附試驗(yan)（ELISA）。往(wang) 預先(xian)包被尿(niao)膽素(su)(su)(su)(su)（urobilin）抗(kang)(kang)體(ti)的(de)(de)(de)(de)(de)包被微(wei)孔(kong)(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)(zhong)，依次(ci)加(jia)(jia)入(ru)(ru)標(biao)(biao)(biao)(biao)(biao)本(ben)(ben)(ben)(ben)、 標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)(pin)(pin)(pin)、HRP標(biao)(biao)(biao)(biao)(biao)記的(de)(de)(de)(de)(de)檢(jian)(jian)測(ce)(ce)抗(kang)(kang)體(ti)，經過(guo)溫(wen)(wen)(wen)育并徹底(di)洗(xi)(xi)(xi)滌。用(yong)(yong)(yong)(yong)(yong)(yong)底(di)物TMB顯 色，TMB在(zai)過(guo)氧化(hua)物酶的(de)(de)(de)(de)(de)催化(hua)下(xia)(xia)轉化(hua)成藍色，并在(zai)酸(suan)的(de)(de)(de)(de)(de)作(zuo)(zuo)用(yong)(yong)(yong)(yong)(yong)(yong)下(xia)(xia)轉化(hua)成 ＊終(zhong)的(de)(de)(de)(de)(de)黃(huang)色。顏色的(de)(de)(de)(de)(de)深淺和樣(yang)品(pin)(pin)(pin)(pin)(pin)(pin)(pin)(pin)中(zhong)(zhong)(zhong)的(de)(de)(de)(de)(de)尿(niao)膽素(su)(su)(su)(su)（urobilin）呈正相關。 用(yong)(yong)(yong)(yong)(yong)(yong)酶標(biao)(biao)(biao)(biao)(biao)儀(yi)在(zai)450nm 波(bo)長下(xia)(xia)測(ce)(ce)定吸光度(du)（OD 值(zhi)），計算樣(yang)品(pin)(pin)(pin)(pin)(pin)(pin)(pin)(pin)濃(nong)(nong)(nong)度(du)。樣(yang)品(pin)(pin)(pin)(pin)(pin)(pin)(pin)(pin)收集、處理(li)及(ji)(ji)保存(cun)方(fang)法1. 血(xue)(xue)(xue)清(qing)(qing)(qing)：使(shi)(shi)用(yong)(yong)(yong)(yong)(yong)(yong)不(bu)(bu)含熱(re)原和內(nei)毒素(su)(su)(su)(su)的(de)(de)(de)(de)(de)試管(guan)，操(cao)(cao)作(zuo)(zuo)過(guo)程中(zhong)(zhong)(zhong)避(bi)(bi)免(mian)任何細(xi) 胞刺激(ji)，收集血(xue)(xue)(xue)液(ye)(ye)(ye)(ye)后(hou)(hou)，3000 轉離心(xin)(xin) 10 分鐘(zhong)將(jiang)血(xue)(xue)(xue)清(qing)(qing)(qing)和紅細(xi)胞迅(xun)速小(xiao)心(xin)(xin) 地(di)分離。 2. 血(xue)(xue)(xue)漿：EDTA、檸檬酸(suan)鹽或(huo)(huo)肝素(su)(su)(su)(su)抗(kang)(kang)凝。3000 轉離心(xin)(xin) 30 分鐘(zhong)取(qu)(qu)上(shang)清(qing)(qing)(qing)。3. 細(xi)胞上(shang)清(qing)(qing)(qing)液(ye)(ye)(ye)(ye)：3000 轉離心(xin)(xin) 10 分鐘(zhong)去(qu)除(chu)顆粒和聚合物。 4. 組織勻(yun)漿：將(jiang)組織加(jia)(jia)入(ru)(ru)適量生理(li)鹽水(shui)搗碎。3000 轉離心(xin)(xin) 10 分鐘(zhong) 取(qu)(qu)上(shang)清(qing)(qing)(qing)。 5. 保存(cun)：如(ru)果(guo)樣(yang)本(ben)(ben)(ben)(ben)收集后(hou)(hou)不(bu)(bu)及(ji)(ji)時(shi)檢(jian)(jian)測(ce)(ce)，請按一次(ci)用(yong)(yong)(yong)(yong)(yong)(yong)量分裝，凍(dong)存(cun) 于(yu)-20℃，避(bi)(bi)免(mian)反(fan)復(fu)(fu)凍(dong)融，在(zai)室(shi)溫(wen)(wen)(wen)下(xia)(xia)解凍(dong)并確(que)保樣(yang)品(pin)(pin)(pin)(pin)(pin)(pin)(pin)(pin)均勻(yun)地(di)充分解凍(dong)。 自備物品(pin)(pin)(pin)(pin)(pin)(pin)(pin)(pin) 1. 酶標(biao)(biao)(biao)(biao)(biao)儀(yi)（450nm）2. 高精度(du)加(jia)(jia)樣(yang)器及(ji)(ji)槍頭：0.5-10uL、2-20uL、20-200uL、200-1000uL 3. 37℃恒(heng)溫(wen)(wen)(wen)箱(xiang) 操(cao)(cao)作(zuo)(zuo)注意事項(xiang) 1. 試劑(ji)盒(he)(he)(he)(he)保存(cun)在(zai) 2-8℃，使(shi)(shi)用(yong)(yong)(yong)(yong)(yong)(yong)前室(shi)溫(wen)(wen)(wen)平衡(heng) 20 分鐘(zhong)。從冰箱(xiang)取(qu)(qu)出(chu)的(de)(de)(de)(de)(de) 濃(nong)(nong)(nong)縮洗(xi)(xi)(xi)滌液(ye)(ye)(ye)(ye)會(hui)有(you)結(jie)(jie)晶(jing)，這(zhe)屬于(yu)正常現象，水(shui)浴(yu)加(jia)(jia)熱(re)使(shi)(shi)結(jie)(jie)晶(jing)完全溶解 后(hou)(hou)再使(shi)(shi)用(yong)(yong)(yong)(yong)(yong)(yong)。2. 實(shi)驗(yan)中(zhong)(zhong)(zhong)不(bu)(bu)用(yong)(yong)(yong)(yong)(yong)(yong)的(de)(de)(de)(de)(de)板(ban)(ban)條(tiao)應立即放回(hui)自封(feng)(feng)袋中(zhong)(zhong)(zhong)，密封(feng)(feng)（低溫(wen)(wen)(wen)干(gan)燥）保 存(cun)。 3. 濃(nong)(nong)(nong)度(du)為(wei) 0 的(de)(de)(de)(de)(de) S0 號標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)(pin)(pin)(pin)即可(ke)視為(wei)陰性對(dui)(dui)照或(huo)(huo)者(zhe)空(kong)白；按照說(shuo)(shuo)明(ming) 書(shu)操(cao)(cao)作(zuo)(zuo)時(shi)樣(yang)本(ben)(ben)(ben)(ben)已經稀釋(shi) 5 倍(bei)(bei)，＊終(zhong)結(jie)(jie)果(guo)乘以 5 才(cai)是樣(yang)本(ben)(ben)(ben)(ben)實(shi)際濃(nong)(nong)(nong)度(du)。4. 嚴格按照說(shuo)(shuo)明(ming)書(shu)中(zhong)(zhong)(zhong)標(biao)(biao)(biao)(biao)(biao)明(ming)的(de)(de)(de)(de)(de)時(shi)間、加(jia)(jia)液(ye)(ye)(ye)(ye)量及(ji)(ji)順(shun)序進行溫(wen)(wen)(wen)育操(cao)(cao)作(zuo)(zuo)。 5. 所有(you)液(ye)(ye)(ye)(ye)體(ti)組分使(shi)(shi)用(yong)(yong)(yong)(yong)(yong)(yong)前充分搖勻(yun)。試劑(ji)盒(he)(he)(he)(he)組成96孔(kong)(kong)(kong)(kong)(kong)(kong)配置48孔(kong)(kong)(kong)(kong)(kong)(kong)配置備注微(wei)孔(kong)(kong)(kong)(kong)(kong)(kong)酶標(biao)(biao)(biao)(biao)(biao)版(ban)12孔(kong)(kong)(kong)(kong)(kong)(kong)*8條(tiao)12孔(kong)(kong)(kong)(kong)(kong)(kong)*4條(tiao)無(wu)標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)(pin)(pin)(pin)0.3ml*6管(guan)0.3ml*6管(guan)無(wu)樣(yang)本(ben)(ben)(ben)(ben)稀釋(shi)液(ye)(ye)(ye)(ye)6ml3ml無(wu)檢(jian)(jian)測(ce)(ce)抗(kang)(kang)體(ti)-HRP10ml5ml無(wu)20*洗(xi)(xi)(xi)滌緩(huan)(huan)(huan)沖(chong)液(ye)(ye)(ye)(ye)25ml15ml按說(shuo)(shuo)明(ming)書(shu)進行稀釋(shi)底(di)物A6ml3ml無(wu)底(di)物B6ml3ml無(wu)終(zhong)止(zhi)液(ye)(ye)(ye)(ye)6ml3ml無(wu)封(feng)(feng)板(ban)(ban)膜2張(zhang)2張(zhang)無(wu)說(shuo)(shuo)明(ming)書(shu)1份1份無(wu)自封(feng)(feng)袋1個1個無(wu)注：標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)(pin)(pin)(pin)（S0-S5）濃(nong)(nong)(nong)度(du)依次(ci)為(wei)：0、10、20、40、80、160 ng/ml 試劑(ji)的(de)(de)(de)(de)(de)準(zhun)(zhun)(zhun)(zhun)備 20×洗(xi)(xi)(xi)滌緩(huan)(huan)(huan)沖(chong)液(ye)(ye)(ye)(ye)的(de)(de)(de)(de)(de)稀釋(shi)：蒸餾(liu)(liu)水(shui)按 1：20 稀釋(shi)，即 1 份的(de)(de)(de)(de)(de) 20×洗(xi)(xi)(xi)滌 緩(huan)(huan)(huan)沖(chong)液(ye)(ye)(ye)(ye)加(jia)(jia) 19 份的(de)(de)(de)(de)(de)蒸餾(liu)(liu)水(shui)。 洗(xi)(xi)(xi)板(ban)(ban)方(fang)法 1. 手(shou)工洗(xi)(xi)(xi)板(ban)(ban)：甩(shuai)盡孔(kong)(kong)(kong)(kong)(kong)(kong)內(nei)液(ye)(ye)(ye)(ye)體(ti)，每(mei)孔(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)滿洗(xi)(xi)(xi)滌液(ye)(ye)(ye)(ye)，靜置 1min 后(hou)(hou)甩(shuai)盡 孔(kong)(kong)(kong)(kong)(kong)(kong)內(nei)液(ye)(ye)(ye)(ye)體(ti)，在(zai)吸水(shui)紙(zhi)上(shang)拍干(gan)，如(ru)此(ci)洗(xi)(xi)(xi)板(ban)(ban) 5 次(ci)。 2. 自動(dong)洗(xi)(xi)(xi)板(ban)(ban)機(ji)：每(mei)孔(kong)(kong)(kong)(kong)(kong)(kong)注入(ru)(ru)洗(xi)(xi)(xi)液(ye)(ye)(ye)(ye) 350μL，浸泡(pao) 1min，洗(xi)(xi)(xi)板(ban)(ban) 5 次(ci)。 操(cao)(cao)作(zuo)(zuo)步驟(zou) 1. 從室(shi)溫(wen)(wen)(wen)平衡(heng) 20min 后(hou)(hou)的(de)(de)(de)(de)(de)鋁箔袋中(zhong)(zhong)(zhong)取(qu)(qu)出(chu)所需板(ban)(ban)條(tiao)，剩(sheng)余板(ban)(ban)條(tiao)用(yong)(yong)(yong)(yong)(yong)(yong)自 封(feng)(feng)袋密封(feng)(feng)放回(hui) 4℃。 2. 設(she)置標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)(pin)(pin)(pin)孔(kong)(kong)(kong)(kong)(kong)(kong)和樣(yang)本(ben)(ben)(ben)(ben)孔(kong)(kong)(kong)(kong)(kong)(kong)，標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)(pin)(pin)(pin)孔(kong)(kong)(kong)(kong)(kong)(kong)各(ge)加(jia)(jia)不(bu)(bu)同濃(nong)(nong)(nong)度(du)的(de)(de)(de)(de)(de)標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)(pin)(pin)(pin) 50μ L；3. 樣(yang)本(ben)(ben)(ben)(ben)孔(kong)(kong)(kong)(kong)(kong)(kong)先(xian)加(jia)(jia)待(dai)測(ce)(ce)樣(yang)本(ben)(ben)(ben)(ben) 10μL，再加(jia)(jia)樣(yang)本(ben)(ben)(ben)(ben)稀釋(shi)液(ye)(ye)(ye)(ye) 40μL(即樣(yang)本(ben)(ben)(ben)(ben)稀 釋(shi) 5 倍(bei)(bei))；空(kong)白孔(kong)(kong)(kong)(kong)(kong)(kong)不(bu)(bu)加(jia)(jia)。4. 除(chu)空(kong)白孔(kong)(kong)(kong)(kong)(kong)(kong)外，標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)(pin)(pin)(pin)孔(kong)(kong)(kong)(kong)(kong)(kong)和樣(yang)本(ben)(ben)(ben)(ben)孔(kong)(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)(zhong)每(mei)孔(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)入(ru)(ru)辣根過(guo)氧化(hua)物酶 （HRP）標(biao)(biao)(biao)(biao)(biao)記的(de)(de)(de)(de)(de)檢(jian)(jian)測(ce)(ce)抗(kang)(kang)體(ti) 100μL，用(yong)(yong)(yong)(yong)(yong)(yong)封(feng)(feng)板(ban)(ban)膜封(feng)(feng)住反(fan)應孔(kong)(kong)(kong)(kong)(kong)(kong)，37℃水(shui)浴(yu)鍋 或(huo)(huo)恒(heng)溫(wen)(wen)(wen)箱(xiang)溫(wen)(wen)(wen)育 60min。 5. 棄去(qu)液(ye)(ye)(ye)(ye)體(ti)，吸水(shui)紙(zhi)上(shang)拍干(gan)，每(mei)孔(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)滿洗(xi)(xi)(xi)滌液(ye)(ye)(ye)(ye)，靜置 1min，甩(shuai)去(qu) 洗(xi)(xi)(xi)滌液(ye)(ye)(ye)(ye)，吸水(shui)紙(zhi)上(shang)拍干(gan)，如(ru)此(ci)重復(fu)(fu)洗(xi)(xi)(xi)板(ban)(ban) 5 次(ci)（也可(ke)用(yong)(yong)(yong)(yong)(yong)(yong)洗(xi)(xi)(xi)板(ban)(ban)機(ji)洗(xi)(xi)(xi)板(ban)(ban)）。6. 每(mei)孔(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)入(ru)(ru)底(di)物 A、B 各(ge) 50μL，37℃避(bi)(bi)光孵育 15min。7. 每(mei)孔(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)入(ru)(ru)終(zhong)止(zhi)液(ye)(ye)(ye)(ye) 50μL，15min 內(nei)，在(zai) 450nm 波(bo)長處測(ce)(ce)定各(ge)孔(kong)(kong)(kong)(kong)(kong)(kong)的(de)(de)(de)(de)(de) OD 值(zhi)。結(jie)(jie)果(guo)判斷(duan)繪制標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)曲線(xian)：在(zai) Excel 工作(zuo)(zuo)表中(zhong)(zhong)(zhong)，以標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)(pin)(pin)(pin)濃(nong)(nong)(nong)度(du)作(zuo)(zuo)橫坐標(biao)(biao)(biao)(biao)(biao)，對(dui)(dui)應 OD 值(zhi)作(zuo)(zuo)縱坐標(biao)(biao)(biao)(biao)(biao)，繪制出(chu)標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)(pin)(pin)(pin)線(xian)性回(hui)歸曲線(xian)，按曲線(xian)方(fang)程計算各(ge)樣(yang) 本(ben)(ben)(ben)(ben)濃(nong)(nong)(nong)度(du)值(zhi)。試劑(ji)盒(he)(he)(he)(he)性能 1. 準(zhun)(zhun)(zhun)(zhun)確(que)性：標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)(pin)(pin)(pin)線(xian)性回(hui)歸與(yu)預期(qi)濃(nong)(nong)(nong)度(du)相關系數(shu) R 值(zhi)，大于(yu)等(deng)于(yu) 0.9900。2. 靈敏(min)度(du)：＊低檢(jian)(jian)測(ce)(ce)濃(nong)(nong)(nong)度(du)小(xiao)于(yu) 1.0 ng/ml。 3. 特(te)異(yi)性：不(bu)(bu)與(yu)其它可(ke)溶性結(jie)(jie)構類(lei)似物交叉反(fan)應。4. 重復(fu)(fu)性：板(ban)(ban)內(nei)、板(ban)(ban)間變異(yi)系數(shu)均小(xiao)于(yu) 15%。 5. 貯藏：2-8℃，避(bi)(bi)光防潮保存(cun)。6. 有(you)效期(qi)：6 個月 免(mian)責聲明(ming) 1. 試劑(ji)盒(he)(he)(he)(he)僅供(gong)研(yan)究使(shi)(shi)用(yong)(yong)(yong)(yong)(yong)(yong)，不(bu)(bu)得(de)用(yong)(yong)(yong)(yong)(yong)(yong)于(yu)臨床實(shi)驗(yan)或(huo)(huo)人(ren)(ren)體(ti)實(shi)驗(yan)，否則所 產生的(de)(de)(de)(de)(de)一切后(hou)(hou)果(guo)，由(you)實(shi)驗(yan)者(zhe)承擔(dan)，本(ben)(ben)(ben)(ben)公司概不(bu)(bu)負責。 2. 嚴格按照說(shuo)(shuo)明(ming)書(shu)操(cao)(cao)作(zuo)(zuo)，實(shi)驗(yan)者(zhe)違(wei)反(fan)說(shuo)(shuo)明(ming)書(shu)操(cao)(cao)作(zuo)(zuo)，后(hou)(hou)果(guo)由(you)實(shi)驗(yan)者(zhe)承擔(dan)人(ren)(ren)（Human）尿(niao)膽素(su)(su)(su)(su)（urobilin）ELISA 檢(jian)(jian)測(ce)(ce)試劑(ji)盒(he)(he)(he)(he)FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES. Human urobilin ELISA Kit instruction Intended use This urobilin ELISA kit is intended Laboratory for Research use only and is not for use in diagnostic or therapeutic procedures.The Stop Solution changes the color from blue to yellow and the intensity of the color is measured at 450 nm using a spectrophotometer. In order to measure the concentration of urobilin in the sample, this urobilin ELISA Kit includes a set of calibration standards. The calibration standards are assayed at the same time as the samples and allow the operator to produce a standard curve of Optical Density versus urobilin concentration. The concentration of urobilin in the samples is then determined by comparing the O.D. of the samples to the standard curve. Sample collection and storages Serum - Use a serum separator tube and allow samples to clot for 30 minutes before centrifugation for 10 minutes at approximately 3000×g. Remove serum and assay immediately or aliquot and store samples at -20℃ or -80℃.Avoid repeated freeze-thaw cycles Plasma - Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 30 minutes at 3000×g at 2-8℃ within 30 minutes of collection. Store samples at -20℃or -80℃. Avoid repeated freeze-thaw cycles.Cell culture supernates and other biological fluids - Remove particulates by centrifugation and assay immediately or aliquot and store samples at -20℃or -80℃. Avoid repeated freeze-thaw cycles. Note: The samples shoule be centrifugated dequately and no hemolysis or granule was allowed. Materials required but not supplied 1. Standard microplate reader(450nm) 2. Precision pipettes and Disposable pipette tips. 3. 37 ℃ incubator Precautions1. Do not substitute reagents from one kit to another. Standard, conjugate microplates are matched for optimal performance. Use only the reagents supplied by manufacturer. 2. Do not remove microplate from the storage bag until needed. Unused strips should be stored at 2-8°C in their pouch with the desiccant provided. 3. Mix all reagents before using. Remove all kit reagents from refrigerator and allow them to reach room temperature ( 20-25°C) Materials supplied Name 96 determinations 48 determinations Microelisa stripplate 12*8strips 12*4strips Standard 0.3ml*6tubes 0.3ml*6tubes Sample Diluent 6.0ml 3.0ml HRP-Conjugate reagent 10.0ml 5.0ml 20X Wash solution 25ml 15ml Chromogen Solution A 6.0ml 3.0ml Chromogen Solution B 6.0ml 3.0ml Stop Solution 6.0ml 3.0ml Closure plate membrane 2 2 User manual 1 1 Sealed bags 1 1Note: Standard (S0 → S5) concentration was followed by:0,10,20,40,80,160 ng/ml Reagent preparation 20×wash solution:Dilute with Distilled or deionized water 1:20.Assay procedure1. Prepare all r e a g e n ts before starting assay procedure. It is recommended that all Standards and Samples be added in duplicate to the Microelisa Stripplate.2. Add standard: Set Standard wells, testing sample wells. Add standard 50μl to standard well. 3. Add Sample: Add testing sample 10μl then add 40μl of Sample Diluent to testing sample well; Blank well doesn’t add anyting. 4. Add 100μl of HRP-conjugate reagent to each well, cover with an adhesive strip and incubate for 60 minutes at 37°C. 5. Aspirate each well and wash, repeating the process four times for a total of five washes. Wash by filling each well with Wash Solution (400μl) using a squirt bottle, manifold dispenser or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Washfllygs.com Solution by aspirating or decanting. Invert the plate and blot it against clean paper towels. 6. Add chromogen solution A 50μl and chromogen solution B 50μl to each well. Gently mix and incubate for 15 minutes at 37°C. Protect from light. 7. Add 50μl Stop Solution to each well. The color in the wells should change from blue to yellow. If the color in the wells is green or the color change does not appear uniform, gently tap the plate to ensure thorough mixing. 8. Read the Optical Density (O.D.) at 450 nm using a microtiter plate reader within 15 minutes. Calculation of results1. This standard curve is used to determine the amount in an unknown sample. The standard curve is generated by plotting the average O.D. (450 nm) obtained for each of the six standard concentrations on the vertical (Y) axis versus the corresponding concentration on the horizontal (X) axis.2. First, calculate the mean O.D. value for each standard and sample. All O.D. values, are subtracted by the mean value of the zero standard before result interpretation. Construct the standard curve using graph paper or statistical software.3. To determine the amount in each sample, first locate the O.D. value on the Y-axis and extend a horizontal line to the standard curve. At the point of intersection, draw a vertical line to the X-axis and read the corresponding concentration. 4. Any variation in operator, pipetting and washing technique, incubation time or temperature, and kit age can cause variation in result. Each user should obtain their own standard curve. 5. The sensitivity by this assay is 1.0 ng/ml 6. Standard curvefllygs.com Storage： 2-8℃. validity： six months. FOR RESEARCH USE ONLY; NOT FOR THERAPEUTIC OR DIAGNOSTIC APPLICATIONS! PLEASE READ THROUGH ENTIRE PROCEDURE BEFORE BEGINNING!
? ??人(ren)組織因(yin)(yin)子(zi)(TF)的(de)(de)(de)(de)(de)(de)(de)elisa??試(shi)(shi)劑(ji)(ji)(ji)盒(he)性(xing)(xing)能1. 特(te)異性(xing)(xing)高(gao)(gao)：由(you)(you)于(yu)(yu)(yu)(yu)是抗(kang)(kang)(kang)原(yuan)(yuan)(yuan)(yuan)抗(kang)(kang)(kang)體(ti)(ti)(ti)(ti)(ti)(ti)的(de)(de)(de)(de)(de)(de)(de)特(te)異性(xing)(xing)結(jie)(jie)合(he)(he)(he)，因(yin)(yin)此(ci)試(shi)(shi)驗(yan)的(de)(de)(de)(de)(de)(de)(de)特(te)異性(xing)(xing)很高(gao)(gao)，能夠排除其他物(wu)(wu)(wu)質的(de)(de)(de)(de)(de)(de)(de)干(gan)擾。2. 靈敏(min)度(du)高(gao)(gao)：由(you)(you)于(yu)(yu)(yu)(yu)酶(mei)(mei)的(de)(de)(de)(de)(de)(de)(de)放大(da)作(zuo)用(yong)(yong)，使(shi)得(de)試(shi)(shi)驗(yan)的(de)(de)(de)(de)(de)(de)(de)靈敏(min)度(du)大(da)大(da)提高(gao)(gao)，能夠檢(jian)(jian)(jian)(jian)(jian)(jian)測(ce)(ce)出(chu)微量的(de)(de)(de)(de)(de)(de)(de)抗(kang)(kang)(kang)原(yuan)(yuan)(yuan)(yuan)或(huo)(huo)抗(kang)(kang)(kang)體(ti)(ti)(ti)(ti)(ti)(ti)。3. 操(cao)(cao)作(zuo)簡(jian)便(bian)：ELISA試(shi)(shi)驗(yan)操(cao)(cao)作(zuo)相對簡(jian)單(dan)，容易掌(zhang)握(wo)，適合(he)(he)(he)于(yu)(yu)(yu)(yu)大(da)規模樣(yang)(yang)本(ben)(ben)(ben)(ben)(ben)檢(jian)(jian)(jian)(jian)(jian)(jian)測(ce)(ce)。4. 適用(yong)(yong)范(fan)圍廣：可以(yi)用(yong)(yong)于(yu)(yu)(yu)(yu)檢(jian)(jian)(jian)(jian)(jian)(jian)測(ce)(ce)各種(zhong)抗(kang)(kang)(kang)原(yuan)(yuan)(yuan)(yuan)、抗(kang)(kang)(kang)體(ti)(ti)(ti)(ti)(ti)(ti)和激素等生(sheng)物(wu)(wu)(wu)分(fen)(fen)(fen)子(zi)。???樣(yang)(yang)品(pin)(pin)(pin)(pin)收集(ji)(ji)、處(chu)理(li)及(ji)保(bao)(bao)(bao)(bao)(bao)(bao)存(cun)(cun)(cun)(cun)方法(fa)(fa)1. 血(xue)(xue)(xue)清：使(shi)用(yong)(yong)不含(han)熱原(yuan)(yuan)(yuan)(yuan)和內毒素的(de)(de)(de)(de)(de)(de)(de)試(shi)(shi)管，操(cao)(cao)作(zuo)過(guo)程中(zhong)(zhong)避(bi)免任(ren)何細胞(bao)刺激，收集(ji)(ji)血(xue)(xue)(xue)液(ye)后(hou)，3000 轉(zhuan)離(li)(li)(li)心(xin)(xin)(xin)(xin) 10 分(fen)(fen)(fen)鐘(zhong)將(jiang)血(xue)(xue)(xue)清和紅細胞(bao)迅速(su)小心(xin)(xin)(xin)(xin)地(di)(di)分(fen)(fen)(fen)離(li)(li)(li)。2. 血(xue)(xue)(xue)漿(jiang)：EDTA、檸檬酸(suan)鹽或(huo)(huo)肝素抗(kang)(kang)(kang)凝。3000 轉(zhuan)離(li)(li)(li)心(xin)(xin)(xin)(xin) 30 分(fen)(fen)(fen)鐘(zhong)取上(shang)(shang)清。3. 細胞(bao)上(shang)(shang)清液(ye)：3000 轉(zhuan)離(li)(li)(li)心(xin)(xin)(xin)(xin) 10 分(fen)(fen)(fen)鐘(zhong)去除顆粒和聚合(he)(he)(he)物(wu)(wu)(wu)。4. 組織勻漿(jiang)：將(jiang)組織加(jia)(jia)入(ru)適量生(sheng)理(li)鹽水(shui)搗碎(sui)。3000 轉(zhuan)離(li)(li)(li)心(xin)(xin)(xin)(xin) 10 分(fen)(fen)(fen)鐘(zhong)取上(shang)(shang)清。5. 保(bao)(bao)(bao)(bao)(bao)(bao)存(cun)(cun)(cun)(cun)：如果(guo)樣(yang)(yang)本(ben)(ben)(ben)(ben)(ben)收集(ji)(ji)后(hou)不及(ji)時(shi)(shi)檢(jian)(jian)(jian)(jian)(jian)(jian)測(ce)(ce)，請按(an)一(yi)(yi)次(ci)用(yong)(yong)量分(fen)(fen)(fen)裝，凍(dong)存(cun)(cun)(cun)(cun)于(yu)(yu)(yu)(yu)-20℃，避(bi)免反(fan)(fan)復(fu)凍(dong)融，在(zai)(zai)室(shi)溫下解 凍(dong)并確(que)保(bao)(bao)(bao)(bao)(bao)(bao)樣(yang)(yang)品(pin)(pin)(pin)(pin)均勻地(di)(di)充分(fen)(fen)(fen)解凍(dong)。?實(shi)(shi)(shi)(shi)驗(yan)原(yuan)(yuan)(yuan)(yuan)理(li)? ? ? 試(shi)(shi)劑(ji)(ji)(ji)盒(he)采用(yong)(yong)雙抗(kang)(kang)(kang)體(ti)(ti)(ti)(ti)(ti)(ti)一(yi)(yi)步(bu)夾(jia)心(xin)(xin)(xin)(xin)法(fa)(fa)酶(mei)(mei)聯吸(xi)附試(shi)(shi)驗(yan)（ELISA）。往(wang)預(yu)先包(bao)被相關指標(biao)(biao)(biao)(biao)(biao)的(de)(de)(de)(de)(de)(de)(de)抗(kang)(kang)(kang)體(ti)(ti)(ti)(ti)(ti)(ti)的(de)(de)(de)(de)(de)(de)(de)包(bao)被微孔(kong)中(zhong)(zhong)，依次(ci)加(jia)(jia)入(ru)標(biao)(biao)(biao)(biao)(biao)本(ben)(ben)(ben)(ben)(ben)、標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)(pin)、HRP標(biao)(biao)(biao)(biao)(biao)記的(de)(de)(de)(de)(de)(de)(de)檢(jian)(jian)(jian)(jian)(jian)(jian)測(ce)(ce)抗(kang)(kang)(kang)體(ti)(ti)(ti)(ti)(ti)(ti)，經過(guo)溫育并洗(xi)滌(di)(di)(di)。用(yong)(yong)底物(wu)(wu)(wu)TMB顯色(se)(se)，TMB在(zai)(zai)過(guo)氧(yang)化(hua)物(wu)(wu)(wu)酶(mei)(mei)的(de)(de)(de)(de)(de)(de)(de)催化(hua)下轉(zhuan)化(hua)成(cheng)(cheng)(cheng)藍色(se)(se)，并在(zai)(zai)酸(suan)的(de)(de)(de)(de)(de)(de)(de)作(zuo)用(yong)(yong)下轉(zhuan)化(hua)成(cheng)(cheng)(cheng)終(zhong)(zhong)的(de)(de)(de)(de)(de)(de)(de)黃(huang)色(se)(se)。顏色(se)(se)的(de)(de)(de)(de)(de)(de)(de)深淺和樣(yang)(yang)品(pin)(pin)(pin)(pin)中(zhong)(zhong)的(de)(de)(de)(de)(de)(de)(de)相關指標(biao)(biao)(biao)(biao)(biao)呈正相關。用(yong)(yong)酶(mei)(mei)標(biao)(biao)(biao)(biao)(biao)儀在(zai)(zai)450nm 波長(chang)(chang)下測(ce)(ce)定吸(xi)光(guang)度(du)（OD 值），計算(suan)樣(yang)(yang)品(pin)(pin)(pin)(pin)濃(nong)(nong)度(du)。??試(shi)(shi)劑(ji)(ji)(ji)盒(he)組成(cheng)(cheng)(cheng)試(shi)(shi)劑(ji)(ji)(ji)盒(he)組成(cheng)(cheng)(cheng)48孔(kong)配置(zhi)96孔(kong)配置(zhi)保(bao)(bao)(bao)(bao)(bao)(bao)存(cun)(cun)(cun)(cun)說(shuo)(shuo)明(ming)(ming)(ming)書1份1份封(feng)(feng)板(ban)膜2片（48）2片（96）密封(feng)(feng)袋(dai)1個1個酶(mei)(mei)標(biao)(biao)(biao)(biao)(biao)包(bao)被板(ban)1×481×962-8℃保(bao)(bao)(bao)(bao)(bao)(bao)存(cun)(cun)(cun)(cun)標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)(pin)0.5ml×1瓶(ping)0.5ml×1瓶(ping)2-8℃保(bao)(bao)(bao)(bao)(bao)(bao)存(cun)(cun)(cun)(cun)標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)(pin)稀釋液(ye)1.5ml×1瓶(ping)1.5ml×1瓶(ping)2-8℃保(bao)(bao)(bao)(bao)(bao)(bao)存(cun)(cun)(cun)(cun)酶(mei)(mei)標(biao)(biao)(biao)(biao)(biao)試(shi)(shi)劑(ji)(ji)(ji)3 ml×1瓶(ping)6 ml×1瓶(ping)2-8℃保(bao)(bao)(bao)(bao)(bao)(bao)存(cun)(cun)(cun)(cun)樣(yang)(yang)品(pin)(pin)(pin)(pin)稀釋液(ye)3 ml×1瓶(ping)6 ml×1瓶(ping)2-8℃保(bao)(bao)(bao)(bao)(bao)(bao)存(cun)(cun)(cun)(cun)顯色(se)(se)劑(ji)(ji)(ji)A液(ye)3 ml×1瓶(ping)6 ml×1瓶(ping)2-8℃保(bao)(bao)(bao)(bao)(bao)(bao)存(cun)(cun)(cun)(cun)顯色(se)(se)劑(ji)(ji)(ji)B液(ye)3 ml×1瓶(ping)6 ml×1瓶(ping)2-8℃保(bao)(bao)(bao)(bao)(bao)(bao)存(cun)(cun)(cun)(cun)終(zhong)(zhong)止(zhi)液(ye)3ml×1瓶(ping)6ml×1瓶(ping)2-8℃保(bao)(bao)(bao)(bao)(bao)(bao)存(cun)(cun)(cun)(cun)20倍濃(nong)(nong)縮洗(xi)滌(di)(di)(di)液(ye)20ml×1瓶(ping)23ml×1瓶(ping)2-8℃保(bao)(bao)(bao)(bao)(bao)(bao)存(cun)(cun)(cun)(cun)??操(cao)(cao)作(zuo)步(bu)驟?1)涂(tu)層:微孔(kong)板(ban)的(de)(de)(de)(de)(de)(de)(de)孔(kong)涂(tu)上(shang)(shang)捕獲抗(kang)(kang)(kang)體(ti)(ti)(ti)(ti)(ti)(ti)或(huo)(huo)抗(kang)(kang)(kang)原(yuan)(yuan)(yuan)(yuan)。2)封(feng)(feng)閉:孔(kong)內加(jia)(jia)入(ru)封(feng)(feng)閉劑(ji)(ji)(ji)(如牛(niu)血(xue)(xue)(xue)清白(bai)蛋白(bai)或(huo)(huo)羧甲基纖維素)，阻止(zhi)非特(te)異性(xing)(xing)蛋白(bai)質結(jie)(jie)合(he)(he)(he)。3)樣(yang)(yang)本(ben)(ben)(ben)(ben)(ben)添(tian)(tian)加(jia)(jia):加(jia)(jia)入(ru)含(han)有未(wei)(wei)知濃(nong)(nong)度(du)的(de)(de)(de)(de)(de)(de)(de)目標(biao)(biao)(biao)(biao)(biao)抗(kang)(kang)(kang)原(yuan)(yuan)(yuan)(yuan)或(huo)(huo)抗(kang)(kang)(kang)體(ti)(ti)(ti)(ti)(ti)(ti)的(de)(de)(de)(de)(de)(de)(de)樣(yang)(yang)本(ben)(ben)(ben)(ben)(ben)。4)洗(xi)滌(di)(di)(di):去除未(wei)(wei)結(jie)(jie)合(he)(he)(he)的(de)(de)(de)(de)(de)(de)(de)物(wu)(wu)(wu)質。5)檢(jian)(jian)(jian)(jian)(jian)(jian)測(ce)(ce)抗(kang)(kang)(kang)體(ti)(ti)(ti)(ti)(ti)(ti)添(tian)(tian)加(jia)(jia):加(jia)(jia)入(ru)對特(te)定抗(kang)(kang)(kang)原(yuan)(yuan)(yuan)(yuan)有特(te)異性(xing)(xing)的(de)(de)(de)(de)(de)(de)(de)酶(mei)(mei)標(biao)(biao)(biao)(biao)(biao)記檢(jian)(jian)(jian)(jian)(jian)(jian)測(ce)(ce)抗(kang)(kang)(kang)體(ti)(ti)(ti)(ti)(ti)(ti)。6) 再次(ci)洗(xi)滌(di)(di)(di):再次(ci)去除未(wei)(wei)結(jie)(jie)合(he)(he)(he)的(de)(de)(de)(de)(de)(de)(de)檢(jian)(jian)(jian)(jian)(jian)(jian)測(ce)(ce)抗(kang)(kang)(kang)體(ti)(ti)(ti)(ti)(ti)(ti)。7)底物(wu)(wu)(wu)添(tian)(tian)加(jia)(jia):加(jia)(jia)入(ru)酶(mei)(mei)的(de)(de)(de)(de)(de)(de)(de)底物(wu)(wu)(wu)，酶(mei)(mei)作(zuo)用(yong)(yong)于(yu)(yu)(yu)(yu)底物(wu)(wu)(wu)產生(sheng)顏色(se)(se)變化(hua)。8)終(zhong)(zhong)止(zhi)反(fan)(fan)應(ying):加(jia)(jia)入(ru)終(zhong)(zhong)止(zhi)溶液(ye)停止(zhi)酶(mei)(mei)的(de)(de)(de)(de)(de)(de)(de)反(fan)(fan)應(ying)(在(zai)(zai)某些類型的(de)(de)(de)(de)(de)(de)(de)ELISA中(zhong)(zhong)需要)。9) 讀數:使(shi)用(yong)(yong)光(guang)譜(pu)光(guang)度(du)計測(ce)(ce)定每個孔(kong)的(de)(de)(de)(de)(de)(de)(de)吸(xi)光(guang)度(du)(通常是在(zai)(zai)特(te)定波長(chang)(chang)下)，吸(xi)光(guang)度(du)與樣(yang)(yang)本(ben)(ben)(ben)(ben)(ben)中(zhong)(zhong)抗(kang)(kang)(kang)原(yuan)(yuan)(yuan)(yuan)或(huo)(huo)抗(kang)(kang)(kang)體(ti)(ti)(ti)(ti)(ti)(ti)的(de)(de)(de)(de)(de)(de)(de)濃(nong)(nong)度(du)成(cheng)(cheng)(cheng)正比。?標(biao)(biao)(biao)(biao)(biao)本(ben)(ben)(ben)(ben)(ben)的(de)(de)(de)(de)(de)(de)(de)采集(ji)(ji)及(ji)保(bao)(bao)(bao)(bao)(bao)(bao)存(cun)(cun)(cun)(cun)1.血(xue)(xue)(xue)清：全血(xue)(xue)(xue)標(biao)(biao)(biao)(biao)(biao)本(ben)(ben)(ben)(ben)(ben)請于(yu)(yu)(yu)(yu)室(shi)溫放置(zhi)2小時(shi)(shi)或(huo)(huo)4℃過(guo)一(yi)(yi)晚后(hou)于(yu)(yu)(yu)(yu)1000 x g離(li)(li)(li)心(xin)(xin)(xin)(xin)20分(fen)(fen)(fen)鐘(zhong)，取上(shang)(shang)清即可檢(jian)(jian)(jian)(jian)(jian)(jian)測(ce)(ce)，或(huo)(huo)將(jiang)標(biao)(biao)(biao)(biao)(biao)本(ben)(ben)(ben)(ben)(ben)放于(yu)(yu)(yu)(yu)-20℃或(huo)(huo)-80℃保(bao)(bao)(bao)(bao)(bao)(bao)存(cun)(cun)(cun)(cun)，但應(ying)避(bi)免反(fan)(fan)復(fu)凍(dong)融。?2.血(xue)(xue)(xue)漿(jiang)：可用(yong)(yong)EDTA或(huo)(huo)肝素作(zuo)為(wei)抗(kang)(kang)(kang)凝劑(ji)(ji)(ji)，標(biao)(biao)(biao)(biao)(biao)本(ben)(ben)(ben)(ben)(ben)采集(ji)(ji)后(hou)30分(fen)(fen)(fen)鐘(zhong)內于(yu)(yu)(yu)(yu)2 - 8° C 1000 x g離(li)(li)(li)心(xin)(xin)(xin)(xin)15分(fen)(fen)(fen)鐘(zhong)，或(huo)(huo)將(jiang)標(biao)(biao)(biao)(biao)(biao)本(ben)(ben)(ben)(ben)(ben)放于(yu)(yu)(yu)(yu)-20℃或(huo)(huo)-80℃保(bao)(bao)(bao)(bao)(bao)(bao)存(cun)(cun)(cun)(cun)，但應(ying)避(bi)免反(fan)(fan)復(fu)凍(dong)融。?3.細胞(bao)培養物(wu)(wu)(wu)上(shang)(shang)清或(huo)(huo)其它生(sheng)物(wu)(wu)(wu)標(biao)(biao)(biao)(biao)(biao)本(ben)(ben)(ben)(ben)(ben)：1000 x g離(li)(li)(li)心(xin)(xin)(xin)(xin)20分(fen)(fen)(fen)鐘(zhong)，取上(shang)(shang)清即可檢(jian)(jian)(jian)(jian)(jian)(jian)測(ce)(ce)，或(huo)(huo)將(jiang)標(biao)(biao)(biao)(biao)(biao)本(ben)(ben)(ben)(ben)(ben)放于(yu)(yu)(yu)(yu)-20℃或(huo)(huo)-80℃保(bao)(bao)(bao)(bao)(bao)(bao)存(cun)(cun)(cun)(cun)，但應(ying)避(bi)免反(fan)(fan)復(fu)凍(dong)融。 (注：標(biao)(biao)(biao)(biao)(biao)本(ben)(ben)(ben)(ben)(ben)溶血(xue)(xue)(xue)會影響(xiang)**檢(jian)(jian)(jian)(jian)(jian)(jian)測(ce)(ce)結(jie)(jie)果(guo)，因(yin)(yin)此(ci)溶血(xue)(xue)(xue)標(biao)(biao)(biao)(biao)(biao)本(ben)(ben)(ben)(ben)(ben)不宜(yi)進行此(ci)項檢(jian)(jian)(jian)(jian)(jian)(jian)測(ce)(ce)。)???結(jie)(jie)果(guo)分(fen)(fen)(fen)析根據試(shi)(shi)劑(ji)(ji)(ji)盒(he)提供(gong)的(de)(de)(de)(de)(de)(de)(de)標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)(pin)濃(nong)(nong)度(du)及(ji)對應(ying)的(de)(de)(de)(de)(de)(de)(de)OD值，利(li)用(yong)(yong)標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)(pin)繪制的(de)(de)(de)(de)(de)(de)(de)標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)曲(qu)線，計算(suan)待測(ce)(ce)樣(yang)(yang)品(pin)(pin)(pin)(pin)的(de)(de)(de)(de)(de)(de)(de)濃(nong)(nong)度(du)。一(yi)(yi)般采用(yong)(yong)擬(ni)合(he)(he)(he)曲(qu)線法(fa)(fa)或(huo)(huo)直線回(hui)歸法(fa)(fa)進行數據處(chu)理(li)，通過(guo)計算(suan)得(de)到待測(ce)(ce)樣(yang)(yang)品(pin)(pin)(pin)(pin)的(de)(de)(de)(de)(de)(de)(de)濃(nong)(nong)度(du)。???免責聲明(ming)(ming)(ming)試(shi)(shi)劑(ji)(ji)(ji)盒(he)僅供(gong)研究使(shi)用(yong)(yong)，不得(de)用(yong)(yong)于(yu)(yu)(yu)(yu)臨床(chuang)實(shi)(shi)(shi)(shi)驗(yan)或(huo)(huo)生(sheng)物(wu)(wu)(wu)體(ti)(ti)(ti)(ti)(ti)(ti)實(shi)(shi)(shi)(shi)驗(yan)，否則所產生(sheng)的(de)(de)(de)(de)(de)(de)(de)一(yi)(yi)切后(hou)果(guo)，由(you)(you)實(shi)(shi)(shi)(shi)驗(yan)者承擔(dan)， 本(ben)(ben)(ben)(ben)(ben)公(gong)司(si)概不負責。嚴格按(an)照說(shuo)(shuo)明(ming)(ming)(ming)書操(cao)(cao)作(zuo)，不同批號不可交換使(shi)用(yong)(yong)，實(shi)(shi)(shi)(shi)驗(yan)者違反(fan)(fan)說(shuo)(shuo)明(ming)(ming)(ming)書操(cao)(cao)作(zuo)，后(hou)果(guo)由(you)(you)實(shi)(shi)(shi)(shi)驗(yan)者承擔(dan)。?
? ??人(ren)二磷酸(suan)(suan)腺苷(gan)(gan)（ADP）ELISA檢(jian)(jian)(jian)測(ce)(ce)試(shi)劑(ji)盒??試(shi)劑(ji)盒性能1. 特異性高(gao)：由于(yu)是抗(kang)(kang)(kang)(kang)(kang)原抗(kang)(kang)(kang)(kang)(kang)體(ti)(ti)(ti)的(de)(de)(de)(de)(de)(de)(de)(de)特異性結(jie)(jie)合(he)(he)(he)，因此(ci)試(shi)驗的(de)(de)(de)(de)(de)(de)(de)(de)特異性很高(gao)，能夠(gou)(gou)排除(chu)其他物(wu)(wu)質(zhi)的(de)(de)(de)(de)(de)(de)(de)(de)干(gan)擾。2. 靈敏度(du)(du)高(gao)：由于(yu)酶的(de)(de)(de)(de)(de)(de)(de)(de)放(fang)大作(zuo)用(yong)(yong)，使(shi)(shi)(shi)得試(shi)驗的(de)(de)(de)(de)(de)(de)(de)(de)靈敏度(du)(du)大大提(ti)高(gao)，能夠(gou)(gou)檢(jian)(jian)(jian)測(ce)(ce)出(chu)微量(liang)的(de)(de)(de)(de)(de)(de)(de)(de)抗(kang)(kang)(kang)(kang)(kang)原或(huo)(huo)(huo)(huo)抗(kang)(kang)(kang)(kang)(kang)體(ti)(ti)(ti)。3. 操作(zuo)簡便：ELISA試(shi)驗操作(zuo)相對簡單，容易掌握，適合(he)(he)(he)于(yu)大規模樣(yang)本(ben)檢(jian)(jian)(jian)測(ce)(ce)。4. 適用(yong)(yong)范圍廣：可以用(yong)(yong)于(yu)檢(jian)(jian)(jian)測(ce)(ce)各種(zhong)抗(kang)(kang)(kang)(kang)(kang)原、抗(kang)(kang)(kang)(kang)(kang)體(ti)(ti)(ti)和(he)激素(su)(su)(su)等(deng)生(sheng)物(wu)(wu)分(fen)(fen)子(zi)。???樣(yang)品收(shou)集(ji)(ji)、處理(li)(li)及(ji)保(bao)存(cun)(cun)方法1. 血(xue)(xue)清：使(shi)(shi)(shi)用(yong)(yong)不(bu)含(han)熱(re)原和(he)內(nei)毒素(su)(su)(su)的(de)(de)(de)(de)(de)(de)(de)(de)試(shi)管，操作(zuo)過(guo)程中(zhong)(zhong)避免(mian)任何細(xi)(xi)胞(bao)刺激，收(shou)集(ji)(ji)血(xue)(xue)液后(hou)，3000 轉離(li)(li)(li)心(xin) 10 分(fen)(fen)鐘(zhong)將(jiang)(jiang)血(xue)(xue)清和(he)紅(hong)細(xi)(xi)胞(bao)迅速小心(xin)地分(fen)(fen)離(li)(li)(li)。2. 血(xue)(xue)漿(jiang)(jiang)：EDTA、檸檬(meng)酸(suan)(suan)鹽或(huo)(huo)(huo)(huo)肝素(su)(su)(su)抗(kang)(kang)(kang)(kang)(kang)凝。3000 轉離(li)(li)(li)心(xin) 30 分(fen)(fen)鐘(zhong)取(qu)(qu)上(shang)(shang)(shang)清。3. 細(xi)(xi)胞(bao)上(shang)(shang)(shang)清液：3000 轉離(li)(li)(li)心(xin) 10 分(fen)(fen)鐘(zhong)去(qu)除(chu)顆(ke)粒和(he)聚(ju)合(he)(he)(he)物(wu)(wu)。4. 組織勻漿(jiang)(jiang)：將(jiang)(jiang)組織加(jia)(jia)入(ru)(ru)(ru)適量(liang)生(sheng)理(li)(li)鹽水搗碎。3000 轉離(li)(li)(li)心(xin) 10 分(fen)(fen)鐘(zhong)取(qu)(qu)上(shang)(shang)(shang)清。5. 保(bao)存(cun)(cun)：如果(guo)樣(yang)本(ben)收(shou)集(ji)(ji)后(hou)不(bu)及(ji)時(shi)檢(jian)(jian)(jian)測(ce)(ce)，請按一次用(yong)(yong)量(liang)分(fen)(fen)裝，凍(dong)存(cun)(cun)于(yu)-20℃，避免(mian)反(fan)復凍(dong)融，在(zai)(zai)室溫下(xia)解 凍(dong)并確保(bao)樣(yang)品均勻地充分(fen)(fen)解凍(dong)。?實(shi)驗原理(li)(li)? ? ? 試(shi)劑(ji)盒采用(yong)(yong)雙抗(kang)(kang)(kang)(kang)(kang)體(ti)(ti)(ti)一步夾心(xin)法酶聯(lian)吸(xi)附(fu)試(shi)驗（ELISA）。往(wang)預先(xian)包(bao)被相關指(zhi)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)的(de)(de)(de)(de)(de)(de)(de)(de)抗(kang)(kang)(kang)(kang)(kang)體(ti)(ti)(ti)的(de)(de)(de)(de)(de)(de)(de)(de)包(bao)被微孔中(zhong)(zhong)，依次加(jia)(jia)入(ru)(ru)(ru)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)本(ben)、標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)品、HRP標(biao)(biao)(biao)(biao)(biao)(biao)(biao)記的(de)(de)(de)(de)(de)(de)(de)(de)檢(jian)(jian)(jian)測(ce)(ce)抗(kang)(kang)(kang)(kang)(kang)體(ti)(ti)(ti)，經過(guo)溫育并洗(xi)滌(di)。用(yong)(yong)底物(wu)(wu)TMB顯色，TMB在(zai)(zai)過(guo)氧化(hua)物(wu)(wu)酶的(de)(de)(de)(de)(de)(de)(de)(de)催化(hua)下(xia)轉化(hua)成(cheng)藍色，并在(zai)(zai)酸(suan)(suan)的(de)(de)(de)(de)(de)(de)(de)(de)作(zuo)用(yong)(yong)下(xia)轉化(hua)成(cheng)終的(de)(de)(de)(de)(de)(de)(de)(de)黃色。顏色的(de)(de)(de)(de)(de)(de)(de)(de)深淺和(he)樣(yang)品中(zhong)(zhong)的(de)(de)(de)(de)(de)(de)(de)(de)相關指(zhi)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)呈正(zheng)相關。用(yong)(yong)酶標(biao)(biao)(biao)(biao)(biao)(biao)(biao)儀(yi)在(zai)(zai)450nm 波(bo)長(chang)下(xia)測(ce)(ce)定(ding)吸(xi)光度(du)(du)（OD 值），計算(suan)樣(yang)品濃度(du)(du)。??試(shi)劑(ji)盒組成(cheng)??操作(zuo)步驟?1)涂層:微孔板的(de)(de)(de)(de)(de)(de)(de)(de)孔涂上(shang)(shang)(shang)捕獲(huo)抗(kang)(kang)(kang)(kang)(kang)體(ti)(ti)(ti)或(huo)(huo)(huo)(huo)抗(kang)(kang)(kang)(kang)(kang)原。2)封(feng)閉(bi):孔內(nei)加(jia)(jia)入(ru)(ru)(ru)封(feng)閉(bi)劑(ji)(如牛(niu)血(xue)(xue)清白蛋(dan)白或(huo)(huo)(huo)(huo)羧甲基纖維素(su)(su)(su))，阻(zu)止(zhi)非特異性蛋(dan)白質(zhi)結(jie)(jie)合(he)(he)(he)。3)樣(yang)本(ben)添加(jia)(jia):加(jia)(jia)入(ru)(ru)(ru)含(han)有未知濃度(du)(du)的(de)(de)(de)(de)(de)(de)(de)(de)目標(biao)(biao)(biao)(biao)(biao)(biao)(biao)抗(kang)(kang)(kang)(kang)(kang)原或(huo)(huo)(huo)(huo)抗(kang)(kang)(kang)(kang)(kang)體(ti)(ti)(ti)的(de)(de)(de)(de)(de)(de)(de)(de)樣(yang)本(ben)。4)洗(xi)滌(di):去(qu)除(chu)未結(jie)(jie)合(he)(he)(he)的(de)(de)(de)(de)(de)(de)(de)(de)物(wu)(wu)質(zhi)。5)檢(jian)(jian)(jian)測(ce)(ce)抗(kang)(kang)(kang)(kang)(kang)體(ti)(ti)(ti)添加(jia)(jia):加(jia)(jia)入(ru)(ru)(ru)對特定(ding)抗(kang)(kang)(kang)(kang)(kang)原有特異性的(de)(de)(de)(de)(de)(de)(de)(de)酶標(biao)(biao)(biao)(biao)(biao)(biao)(biao)記檢(jian)(jian)(jian)測(ce)(ce)抗(kang)(kang)(kang)(kang)(kang)體(ti)(ti)(ti)。6) 再次洗(xi)滌(di):再次去(qu)除(chu)未結(jie)(jie)合(he)(he)(he)的(de)(de)(de)(de)(de)(de)(de)(de)檢(jian)(jian)(jian)測(ce)(ce)抗(kang)(kang)(kang)(kang)(kang)體(ti)(ti)(ti)。7)底物(wu)(wu)添加(jia)(jia):加(jia)(jia)入(ru)(ru)(ru)酶的(de)(de)(de)(de)(de)(de)(de)(de)底物(wu)(wu)，酶作(zuo)用(yong)(yong)于(yu)底物(wu)(wu)產生(sheng)顏色變化(hua)。8)終止(zhi)反(fan)應(ying):加(jia)(jia)入(ru)(ru)(ru)終止(zhi)溶液停止(zhi)酶的(de)(de)(de)(de)(de)(de)(de)(de)反(fan)應(ying)(在(zai)(zai)某些(xie)類(lei)型的(de)(de)(de)(de)(de)(de)(de)(de)ELISA中(zhong)(zhong)需要)。9) 讀數:使(shi)(shi)(shi)用(yong)(yong)光譜光度(du)(du)計測(ce)(ce)定(ding)每個孔的(de)(de)(de)(de)(de)(de)(de)(de)吸(xi)光度(du)(du)(通常是在(zai)(zai)特定(ding)波(bo)長(chang)下(xia))，吸(xi)光度(du)(du)與樣(yang)本(ben)中(zhong)(zhong)抗(kang)(kang)(kang)(kang)(kang)原或(huo)(huo)(huo)(huo)抗(kang)(kang)(kang)(kang)(kang)體(ti)(ti)(ti)的(de)(de)(de)(de)(de)(de)(de)(de)濃度(du)(du)成(cheng)正(zheng)比。?人(ren)二磷酸(suan)(suan)腺苷(gan)(gan)（ADP）ELISA檢(jian)(jian)(jian)測(ce)(ce)試(shi)劑(ji)盒標(biao)(biao)(biao)(biao)(biao)(biao)(biao)本(ben)的(de)(de)(de)(de)(de)(de)(de)(de)采集(ji)(ji)及(ji)保(bao)存(cun)(cun)1.血(xue)(xue)清：全血(xue)(xue)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)本(ben)請于(yu)室溫放(fang)置2小時(shi)或(huo)(huo)(huo)(huo)4℃過(guo)一晚后(hou)于(yu)1000 x g離(li)(li)(li)心(xin)20分(fen)(fen)鐘(zhong)，取(qu)(qu)上(shang)(shang)(shang)清即可檢(jian)(jian)(jian)測(ce)(ce)，或(huo)(huo)(huo)(huo)將(jiang)(jiang)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)本(ben)放(fang)于(yu)-20℃或(huo)(huo)(huo)(huo)-80℃保(bao)存(cun)(cun)，但(dan)應(ying)避免(mian)反(fan)復凍(dong)融。?2.血(xue)(xue)漿(jiang)(jiang)：可用(yong)(yong)EDTA或(huo)(huo)(huo)(huo)肝素(su)(su)(su)作(zuo)為(wei)抗(kang)(kang)(kang)(kang)(kang)凝劑(ji)，標(biao)(biao)(biao)(biao)(biao)(biao)(biao)本(ben)采集(ji)(ji)后(hou)30分(fen)(fen)鐘(zhong)內(nei)于(yu)2 - 8° C 1000 x g離(li)(li)(li)心(xin)15分(fen)(fen)鐘(zhong)，或(huo)(huo)(huo)(huo)將(jiang)(jiang)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)本(ben)放(fang)于(yu)-20℃或(huo)(huo)(huo)(huo)-80℃保(bao)存(cun)(cun)，但(dan)應(ying)避免(mian)反(fan)復凍(dong)融。?3.細(xi)(xi)胞(bao)培養物(wu)(wu)上(shang)(shang)(shang)清或(huo)(huo)(huo)(huo)其它生(sheng)物(wu)(wu)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)本(ben)：1000 x g離(li)(li)(li)心(xin)20分(fen)(fen)鐘(zhong)，取(qu)(qu)上(shang)(shang)(shang)清即可檢(jian)(jian)(jian)測(ce)(ce)，或(huo)(huo)(huo)(huo)將(jiang)(jiang)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)本(ben)放(fang)于(yu)-20℃或(huo)(huo)(huo)(huo)-80℃保(bao)存(cun)(cun)，但(dan)應(ying)避免(mian)反(fan)復凍(dong)融。 (注(zhu)：標(biao)(biao)(biao)(biao)(biao)(biao)(biao)本(ben)溶血(xue)(xue)會(hui)影響**檢(jian)(jian)(jian)測(ce)(ce)結(jie)(jie)果(guo)，因此(ci)溶血(xue)(xue)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)本(ben)不(bu)宜(yi)進(jin)(jin)行此(ci)項檢(jian)(jian)(jian)測(ce)(ce)。)???結(jie)(jie)果(guo)分(fen)(fen)析根據(ju)試(shi)劑(ji)盒提(ti)供(gong)(gong)的(de)(de)(de)(de)(de)(de)(de)(de)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)品濃度(du)(du)及(ji)對應(ying)的(de)(de)(de)(de)(de)(de)(de)(de)OD值，利(li)用(yong)(yong)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)品繪制的(de)(de)(de)(de)(de)(de)(de)(de)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)曲(qu)線，計算(suan)待(dai)(dai)測(ce)(ce)樣(yang)品的(de)(de)(de)(de)(de)(de)(de)(de)濃度(du)(du)。一般采用(yong)(yong)擬合(he)(he)(he)曲(qu)線法或(huo)(huo)(huo)(huo)直線回歸(gui)法進(jin)(jin)行數據(ju)處理(li)(li)，通過(guo)計算(suan)得到待(dai)(dai)測(ce)(ce)樣(yang)品的(de)(de)(de)(de)(de)(de)(de)(de)濃度(du)(du)。???免(mian)責(ze)聲明試(shi)劑(ji)盒僅供(gong)(gong)研究(jiu)使(shi)(shi)(shi)用(yong)(yong)，不(bu)得用(yong)(yong)于(yu)臨床實(shi)驗或(huo)(huo)(huo)(huo)生(sheng)物(wu)(wu)體(ti)(ti)(ti)實(shi)驗，否則(ze)所產生(sheng)的(de)(de)(de)(de)(de)(de)(de)(de)一切后(hou)果(guo)，由實(shi)驗者(zhe)(zhe)承(cheng)擔， 本(ben)公司概不(bu)負責(ze)。嚴格按照說(shuo)明書操作(zuo)，不(bu)同(tong)批(pi)號不(bu)可交(jiao)換使(shi)(shi)(shi)用(yong)(yong)，實(shi)驗者(zhe)(zhe)違反(fan)說(shuo)明書操作(zuo)，后(hou)果(guo)由實(shi)驗者(zhe)(zhe)承(cheng)擔。?
; 人含纈酪(lao)肽蛋(dan)白(bai)（VCP）ELISA檢(jian)(jian)(jian)測試(shi)(shi)劑(ji)盒(he) 實(shi)驗(yan)(yan)原理(li)(li) ELISA運(yun)用(yong)(yong)了免疫(yi)學(xue)(xue)原理(li)(li)和(he)(he)(he)化學(xue)(xue)反應顯色原理(li)(li)，需(xu)要將(jiang)抗(kang)原或(huo)(huo)抗(kang)體(ti)(ti)預包被在(zai)(zai)(zai)固相(xiang)載(zai)體(ti)(ti)表面，使后來形成的(de)(de)抗(kang)原-抗(kang)體(ti)(ti)-酶(mei)(mei) (熒光基團) -底(di)物(wu)(wu)復合物(wu)(wu)黏附在(zai)(zai)(zai)載(zai)體(ti)(ti)上(shang)，通過(guo)(guo)檢(jian)(jian)(jian)測OD值(zhi)或(huo)(huo)發(fa)光值(zhi)，來檢(jian)(jian)(jian)測靶蛋(dan)白(bai)的(de)(de)含量。 實(shi)驗(yan)(yan)所需(xu)自(zi)備(bei)物(wu)(wu)品(pin)(pin)1. 酶(mei)(mei)標(biao)(biao)(biao)儀（450nm）2. 加(jia)(jia)樣(yang)(yang)器及(ji)槍頭：0.5-10uL、2-20uL、20-200uL、200-1000uL3. 37℃恒溫(wen)(wen)箱(xiang)(xiang) 人含纈酪(lao)肽蛋(dan)白(bai)（VCP）ELISA檢(jian)(jian)(jian)測試(shi)(shi)劑(ji)盒(he)試(shi)(shi)劑(ji)盒(he)組(zu)成名稱96 孔(kong)(kong)(kong)配置48 孔(kong)(kong)(kong)配置備(bei)注微孔(kong)(kong)(kong)酶(mei)(mei)標(biao)(biao)(biao)板12 孔(kong)(kong)(kong)×8 條12 孔(kong)(kong)(kong)×4 條無(wu)(wu)標(biao)(biao)(biao)準(zhun)品(pin)(pin)0.5mL0.5mL按說明書(shu)(shu)進行稀(xi)釋標(biao)(biao)(biao)準(zhun)品(pin)(pin)稀(xi)釋液(ye)6mL3mL按說明書(shu)(shu)進行稀(xi)釋樣(yang)(yang)本(ben)稀(xi)釋液(ye)6mL3mL按說明書(shu)(shu)進行稀(xi)釋檢(jian)(jian)(jian)測抗(kang)體(ti)(ti)-HRP6mL3mL無(wu)(wu)20×洗(xi)(xi)滌(di)緩沖液(ye)20mL20mL按說明書(shu)(shu)進行稀(xi)釋底(di)物(wu)(wu) A6mL3mL無(wu)(wu)底(di)物(wu)(wu) B6mL3mL無(wu)(wu)終止(zhi)液(ye)6mL3mL無(wu)(wu)封板膜2 張2 張無(wu)(wu)說明書(shu)(shu)1 份1 份無(wu)(wu)自(zi)封袋1 個(ge)1 個(ge)無(wu)(wu) 操(cao)作(zuo)步驟(zou)1. 準(zhun)備(bei)工(gong)作(zuo)：將(jiang)試(shi)(shi)劑(ji)盒(he)從冰箱(xiang)(xiang)中(zhong)取(qu)(qu)出，室(shi)溫(wen)(wen)放置30分(fen)(fen)鐘，使試(shi)(shi)劑(ji)盒(he)恢復至室(shi)溫(wen)(wen)。同時(shi)準(zhun)備(bei)好實(shi)驗(yan)(yan)所需(xu)的(de)(de)材料和(he)(he)(he)器具。2. 加(jia)(jia)樣(yang)(yang)：在(zai)(zai)(zai)酶(mei)(mei)標(biao)(biao)(biao)板的(de)(de)弟(di)1個(ge)孔(kong)(kong)(kong)中(zhong)加(jia)(jia)入(ru)(ru)標(biao)(biao)(biao)準(zhun)品(pin)(pin)50μL，然(ran)后在(zai)(zai)(zai)后續(xu)的(de)(de)孔(kong)(kong)(kong)中(zhong)依次(ci)加(jia)(jia)入(ru)(ru)待測樣(yang)(yang)本(ben)50μL。3. 溫(wen)(wen)育(yu)：將(jiang)酶(mei)(mei)標(biao)(biao)(biao)板密封，在(zai)(zai)(zai)37℃下溫(wen)(wen)育(yu)30分(fen)(fen)鐘。4. 洗(xi)(xi)滌(di)：用(yong)(yong)洗(xi)(xi)滌(di)液(ye)清洗(xi)(xi)酶(mei)(mei)標(biao)(biao)(biao)板，共(gong)洗(xi)(xi)滌(di)5次(ci)。每次(ci)洗(xi)(xi)滌(di)后，用(yong)(yong)吸(xi)水(shui)紙輕(qing)輕(qing)吸(xi)干孔(kong)(kong)(kong)內(nei)液(ye)體(ti)(ti)。5. 加(jia)(jia)酶(mei)(mei)：在(zai)(zai)(zai)每個(ge)孔(kong)(kong)(kong)中(zhong)加(jia)(jia)入(ru)(ru)酶(mei)(mei)標(biao)(biao)(biao)物(wu)(wu)100μL。6. 溫(wen)(wen)育(yu)：再(zai)次(ci)將(jiang)酶(mei)(mei)標(biao)(biao)(biao)板密封，在(zai)(zai)(zai)37℃下溫(wen)(wen)育(yu)30分(fen)(fen)鐘。7. 洗(xi)(xi)滌(di)：用(yong)(yong)洗(xi)(xi)滌(di)液(ye)清洗(xi)(xi)酶(mei)(mei)標(biao)(biao)(biao)板，共(gong)洗(xi)(xi)滌(di)5次(ci)。每次(ci)洗(xi)(xi)滌(di)后，用(yong)(yong)吸(xi)水(shui)紙輕(qing)輕(qing)吸(xi)干孔(kong)(kong)(kong)內(nei)液(ye)體(ti)(ti)。8. 加(jia)(jia)底(di)物(wu)(wu)：在(zai)(zai)(zai)每個(ge)孔(kong)(kong)(kong)中(zhong)加(jia)(jia)入(ru)(ru)底(di)物(wu)(wu)A液(ye)50μL，再(zai)加(jia)(jia)入(ru)(ru)底(di)物(wu)(wu)B液(ye)50μL。9. 終止(zhi)反應：在(zai)(zai)(zai)每個(ge)孔(kong)(kong)(kong)中(zhong)加(jia)(jia)入(ru)(ru)終止(zhi)液(ye)50μL。10. 檢(jian)(jian)(jian)測：用(yong)(yong)酶(mei)(mei)標(biao)(biao)(biao)儀在(zai)(zai)(zai)450nm波(bo)(bo)長下讀(du)取(qu)(qu)各孔(kong)(kong)(kong)的(de)(de)吸(xi)光度(du)(du)（OD值(zhi)）。 結(jie)果分(fen)(fen)析(xi)根據標(biao)(biao)(biao)準(zhun)品(pin)(pin)和(he)(he)(he)樣(yang)(yang)本(ben)的(de)(de)OD值(zhi)，繪制(zhi)標(biao)(biao)(biao)準(zhun)曲線(xian)，從而計算出樣(yang)(yang)本(ben)中(zhong)角(jiao)蛋(dan)白(bai)1（KRT1）的(de)(de)濃(nong)度(du)(du)。具體(ti)(ti)分(fen)(fen)析(xi)方法可參考(kao)試(shi)(shi)劑(ji)盒(he)說明書(shu)(shu)或(huo)(huo)相(xiang)關(guan)文獻。注意事(shi)項1. 在(zai)(zai)(zai)操(cao)作(zuo)過(guo)(guo)程(cheng)(cheng)中(zhong)，應保(bao)持酶(mei)(mei)標(biao)(biao)(biao)板的(de)(de)干燥，避(bi)免水(shui)分(fen)(fen)進入(ru)(ru)孔(kong)(kong)(kong)內(nei)。2. 加(jia)(jia)樣(yang)(yang)時(shi)需(xu)確(que)保(bao)加(jia)(jia)樣(yang)(yang)準(zhun)確(que)、無(wu)(wu)氣泡產生(sheng)。3. 溫(wen)(wen)育(yu)和(he)(he)(he)洗(xi)(xi)滌(di)過(guo)(guo)程(cheng)(cheng)中(zhong)需(xu)嚴格控制(zhi)時(shi)間(jian)和(he)(he)(he)溫(wen)(wen)度(du)(du)，確(que)保(bao)實(shi)驗(yan)(yan)結(jie)果的(de)(de)準(zhun)確(que)性。4. 在(zai)(zai)(zai)使用(yong)(yong)酶(mei)(mei)標(biao)(biao)(biao)儀讀(du)取(qu)(qu)OD值(zhi)時(shi)，需(xu)確(que)保(bao)波(bo)(bo)長設置正確(que)，避(bi)免誤差產生(sheng)。5. 在(zai)(zai)(zai)整個(ge)實(shi)驗(yan)(yan)過(guo)(guo)程(cheng)(cheng)中(zhong)，需(xu)保(bao)持操(cao)作(zuo)環境的(de)(de)清潔和(he)(he)(he)整潔，避(bi)免交(jiao)叉污染。
? ??人(ren)（Human）載脂(zhi)蛋白 E（apo-E）ELISA 檢(jian)(jian)測(ce)(ce)(ce)試(shi)(shi)(shi)劑(ji)盒??試(shi)(shi)(shi)劑(ji)盒性能1. 特異性高(gao)：由(you)(you)于(yu)(yu)是抗(kang)(kang)(kang)(kang)(kang)原(yuan)抗(kang)(kang)(kang)(kang)(kang)體(ti)(ti)的(de)(de)(de)(de)(de)(de)特異性結(jie)(jie)合(he)，因(yin)(yin)此(ci)試(shi)(shi)(shi)驗(yan)(yan)(yan)(yan)(yan)(yan)的(de)(de)(de)(de)(de)(de)特異性很高(gao)，能夠(gou)排除(chu)其他物(wu)(wu)質的(de)(de)(de)(de)(de)(de)干擾(rao)。2. 靈敏度(du)(du)(du)高(gao)：由(you)(you)于(yu)(yu)酶(mei)(mei)的(de)(de)(de)(de)(de)(de)放大作(zuo)(zuo)(zuo)(zuo)用(yong)(yong)，使(shi)得試(shi)(shi)(shi)驗(yan)(yan)(yan)(yan)(yan)(yan)的(de)(de)(de)(de)(de)(de)靈敏度(du)(du)(du)大大提高(gao)，能夠(gou)檢(jian)(jian)測(ce)(ce)(ce)出微(wei)量(liang)的(de)(de)(de)(de)(de)(de)抗(kang)(kang)(kang)(kang)(kang)原(yuan)或(huo)(huo)(huo)(huo)(huo)(huo)(huo)(huo)抗(kang)(kang)(kang)(kang)(kang)體(ti)(ti)。3. 操(cao)(cao)(cao)作(zuo)(zuo)(zuo)(zuo)簡(jian)便：ELISA試(shi)(shi)(shi)驗(yan)(yan)(yan)(yan)(yan)(yan)操(cao)(cao)(cao)作(zuo)(zuo)(zuo)(zuo)相(xiang)對簡(jian)單，容易掌(zhang)握(wo)，適合(he)于(yu)(yu)大規模樣(yang)(yang)本(ben)(ben)(ben)(ben)檢(jian)(jian)測(ce)(ce)(ce)。4. 適用(yong)(yong)范圍(wei)廣(guang)：可以(yi)用(yong)(yong)于(yu)(yu)檢(jian)(jian)測(ce)(ce)(ce)各(ge)種抗(kang)(kang)(kang)(kang)(kang)原(yuan)、抗(kang)(kang)(kang)(kang)(kang)體(ti)(ti)和(he)(he)(he)激(ji)素(su)等生(sheng)物(wu)(wu)分(fen)(fen)子(zi)。???樣(yang)(yang)品(pin)(pin)(pin)(pin)收集(ji)、處(chu)理及(ji)保(bao)(bao)存(cun)方法(fa)(fa)1. 血(xue)清(qing)(qing)：使(shi)用(yong)(yong)不(bu)(bu)(bu)含熱原(yuan)和(he)(he)(he)內(nei)毒(du)素(su)的(de)(de)(de)(de)(de)(de)試(shi)(shi)(shi)管，操(cao)(cao)(cao)作(zuo)(zuo)(zuo)(zuo)過程中(zhong)避(bi)(bi)免(mian)(mian)任何細(xi)(xi)胞刺(ci)激(ji)，收集(ji)血(xue)液(ye)后(hou)，3000 轉離(li)(li)心 10 分(fen)(fen)鐘(zhong)(zhong)將(jiang)(jiang)血(xue)清(qing)(qing)和(he)(he)(he)紅細(xi)(xi)胞迅(xun)速小(xiao)心地分(fen)(fen)離(li)(li)。2. 血(xue)漿：EDTA、檸(ning)檬酸鹽(yan)或(huo)(huo)(huo)(huo)(huo)(huo)(huo)(huo)肝素(su)抗(kang)(kang)(kang)(kang)(kang)凝。3000 轉離(li)(li)心 30 分(fen)(fen)鐘(zhong)(zhong)取(qu)(qu)上清(qing)(qing)。3. 細(xi)(xi)胞上清(qing)(qing)液(ye)：3000 轉離(li)(li)心 10 分(fen)(fen)鐘(zhong)(zhong)去(qu)除(chu)顆粒和(he)(he)(he)聚(ju)合(he)物(wu)(wu)。4. 組織勻漿：將(jiang)(jiang)組織加(jia)(jia)(jia)入(ru)適量(liang)生(sheng)理鹽(yan)水搗(dao)碎。3000 轉離(li)(li)心 10 分(fen)(fen)鐘(zhong)(zhong)取(qu)(qu)上清(qing)(qing)。5. 保(bao)(bao)存(cun)：如果樣(yang)(yang)本(ben)(ben)(ben)(ben)收集(ji)后(hou)不(bu)(bu)(bu)及(ji)時(shi)檢(jian)(jian)測(ce)(ce)(ce)，請(qing)按一(yi)次用(yong)(yong)量(liang)分(fen)(fen)裝(zhuang)，凍(dong)存(cun)于(yu)(yu)-20℃，避(bi)(bi)免(mian)(mian)反(fan)復(fu)凍(dong)融(rong)，在(zai)(zai)(zai)室溫下(xia)解(jie)(jie) 凍(dong)并確保(bao)(bao)樣(yang)(yang)品(pin)(pin)(pin)(pin)均(jun)勻地充分(fen)(fen)解(jie)(jie)凍(dong)。?實(shi)驗(yan)(yan)(yan)(yan)(yan)(yan)原(yuan)理? ? ? 試(shi)(shi)(shi)劑(ji)盒采(cai)(cai)用(yong)(yong)雙抗(kang)(kang)(kang)(kang)(kang)體(ti)(ti)一(yi)步夾心法(fa)(fa)酶(mei)(mei)聯吸附(fu)試(shi)(shi)(shi)驗(yan)(yan)(yan)(yan)(yan)(yan)（ELISA）。往預先包被相(xiang)關指標(biao)(biao)(biao)(biao)的(de)(de)(de)(de)(de)(de)抗(kang)(kang)(kang)(kang)(kang)體(ti)(ti)的(de)(de)(de)(de)(de)(de)包被微(wei)孔(kong)中(zhong)，依次加(jia)(jia)(jia)入(ru)標(biao)(biao)(biao)(biao)本(ben)(ben)(ben)(ben)、標(biao)(biao)(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)(pin)、HRP標(biao)(biao)(biao)(biao)記的(de)(de)(de)(de)(de)(de)檢(jian)(jian)測(ce)(ce)(ce)抗(kang)(kang)(kang)(kang)(kang)體(ti)(ti)，經過溫育并洗滌。用(yong)(yong)底(di)物(wu)(wu)TMB顯(xian)色，TMB在(zai)(zai)(zai)過氧化(hua)(hua)物(wu)(wu)酶(mei)(mei)的(de)(de)(de)(de)(de)(de)催化(hua)(hua)下(xia)轉化(hua)(hua)成藍色，并在(zai)(zai)(zai)酸的(de)(de)(de)(de)(de)(de)作(zuo)(zuo)(zuo)(zuo)用(yong)(yong)下(xia)轉化(hua)(hua)成終(zhong)的(de)(de)(de)(de)(de)(de)黃色。顏色的(de)(de)(de)(de)(de)(de)深淺(qian)和(he)(he)(he)樣(yang)(yang)品(pin)(pin)(pin)(pin)中(zhong)的(de)(de)(de)(de)(de)(de)相(xiang)關指標(biao)(biao)(biao)(biao)呈正(zheng)相(xiang)關。用(yong)(yong)酶(mei)(mei)標(biao)(biao)(biao)(biao)儀在(zai)(zai)(zai)450nm 波(bo)長下(xia)測(ce)(ce)(ce)定(ding)吸光(guang)(guang)(guang)度(du)(du)(du)（OD 值），計(ji)(ji)算(suan)樣(yang)(yang)品(pin)(pin)(pin)(pin)濃(nong)度(du)(du)(du)。??試(shi)(shi)(shi)劑(ji)盒組成??操(cao)(cao)(cao)作(zuo)(zuo)(zuo)(zuo)步驟?1)涂層:微(wei)孔(kong)板的(de)(de)(de)(de)(de)(de)孔(kong)涂上捕獲抗(kang)(kang)(kang)(kang)(kang)體(ti)(ti)或(huo)(huo)(huo)(huo)(huo)(huo)(huo)(huo)抗(kang)(kang)(kang)(kang)(kang)原(yuan)。2)封閉:孔(kong)內(nei)加(jia)(jia)(jia)入(ru)封閉劑(ji)(如牛血(xue)清(qing)(qing)白蛋白或(huo)(huo)(huo)(huo)(huo)(huo)(huo)(huo)羧甲基纖(xian)維素(su))，阻止(zhi)非特異性蛋白質結(jie)(jie)合(he)。3)樣(yang)(yang)本(ben)(ben)(ben)(ben)添加(jia)(jia)(jia):加(jia)(jia)(jia)入(ru)含有(you)未知濃(nong)度(du)(du)(du)的(de)(de)(de)(de)(de)(de)目標(biao)(biao)(biao)(biao)抗(kang)(kang)(kang)(kang)(kang)原(yuan)或(huo)(huo)(huo)(huo)(huo)(huo)(huo)(huo)抗(kang)(kang)(kang)(kang)(kang)體(ti)(ti)的(de)(de)(de)(de)(de)(de)樣(yang)(yang)本(ben)(ben)(ben)(ben)。4)洗滌:去(qu)除(chu)未結(jie)(jie)合(he)的(de)(de)(de)(de)(de)(de)物(wu)(wu)質。5)檢(jian)(jian)測(ce)(ce)(ce)抗(kang)(kang)(kang)(kang)(kang)體(ti)(ti)添加(jia)(jia)(jia):加(jia)(jia)(jia)入(ru)對特定(ding)抗(kang)(kang)(kang)(kang)(kang)原(yuan)有(you)特異性的(de)(de)(de)(de)(de)(de)酶(mei)(mei)標(biao)(biao)(biao)(biao)記檢(jian)(jian)測(ce)(ce)(ce)抗(kang)(kang)(kang)(kang)(kang)體(ti)(ti)。6) 再次洗滌:再次去(qu)除(chu)未結(jie)(jie)合(he)的(de)(de)(de)(de)(de)(de)檢(jian)(jian)測(ce)(ce)(ce)抗(kang)(kang)(kang)(kang)(kang)體(ti)(ti)。7)底(di)物(wu)(wu)添加(jia)(jia)(jia):加(jia)(jia)(jia)入(ru)酶(mei)(mei)的(de)(de)(de)(de)(de)(de)底(di)物(wu)(wu)，酶(mei)(mei)作(zuo)(zuo)(zuo)(zuo)用(yong)(yong)于(yu)(yu)底(di)物(wu)(wu)產生(sheng)顏色變化(hua)(hua)。8)終(zhong)止(zhi)反(fan)應:加(jia)(jia)(jia)入(ru)終(zhong)止(zhi)溶(rong)液(ye)停止(zhi)酶(mei)(mei)的(de)(de)(de)(de)(de)(de)反(fan)應(在(zai)(zai)(zai)某些(xie)類型的(de)(de)(de)(de)(de)(de)ELISA中(zhong)需要)。9) 讀數:使(shi)用(yong)(yong)光(guang)(guang)(guang)譜光(guang)(guang)(guang)度(du)(du)(du)計(ji)(ji)測(ce)(ce)(ce)定(ding)每個孔(kong)的(de)(de)(de)(de)(de)(de)吸光(guang)(guang)(guang)度(du)(du)(du)(通常是在(zai)(zai)(zai)特定(ding)波(bo)長下(xia))，吸光(guang)(guang)(guang)度(du)(du)(du)與樣(yang)(yang)本(ben)(ben)(ben)(ben)中(zhong)抗(kang)(kang)(kang)(kang)(kang)原(yuan)或(huo)(huo)(huo)(huo)(huo)(huo)(huo)(huo)抗(kang)(kang)(kang)(kang)(kang)體(ti)(ti)的(de)(de)(de)(de)(de)(de)濃(nong)度(du)(du)(du)成正(zheng)比(bi)。?標(biao)(biao)(biao)(biao)本(ben)(ben)(ben)(ben)的(de)(de)(de)(de)(de)(de)采(cai)(cai)集(ji)及(ji)保(bao)(bao)存(cun)1.血(xue)清(qing)(qing)：全(quan)血(xue)標(biao)(biao)(biao)(biao)本(ben)(ben)(ben)(ben)請(qing)于(yu)(yu)室溫放置2小(xiao)時(shi)或(huo)(huo)(huo)(huo)(huo)(huo)(huo)(huo)4℃過一(yi)晚后(hou)于(yu)(yu)1000 x g離(li)(li)心20分(fen)(fen)鐘(zhong)(zhong)，取(qu)(qu)上清(qing)(qing)即(ji)可檢(jian)(jian)測(ce)(ce)(ce)，或(huo)(huo)(huo)(huo)(huo)(huo)(huo)(huo)將(jiang)(jiang)標(biao)(biao)(biao)(biao)本(ben)(ben)(ben)(ben)放于(yu)(yu)-20℃或(huo)(huo)(huo)(huo)(huo)(huo)(huo)(huo)-80℃保(bao)(bao)存(cun)，但(dan)應避(bi)(bi)免(mian)(mian)反(fan)復(fu)凍(dong)融(rong)。?2.血(xue)漿：可用(yong)(yong)EDTA或(huo)(huo)(huo)(huo)(huo)(huo)(huo)(huo)肝素(su)作(zuo)(zuo)(zuo)(zuo)為抗(kang)(kang)(kang)(kang)(kang)凝劑(ji)，標(biao)(biao)(biao)(biao)本(ben)(ben)(ben)(ben)采(cai)(cai)集(ji)后(hou)30分(fen)(fen)鐘(zhong)(zhong)內(nei)于(yu)(yu)2 - 8° C 1000 x g離(li)(li)心15分(fen)(fen)鐘(zhong)(zhong)，或(huo)(huo)(huo)(huo)(huo)(huo)(huo)(huo)將(jiang)(jiang)標(biao)(biao)(biao)(biao)本(ben)(ben)(ben)(ben)放于(yu)(yu)-20℃或(huo)(huo)(huo)(huo)(huo)(huo)(huo)(huo)-80℃保(bao)(bao)存(cun)，但(dan)應避(bi)(bi)免(mian)(mian)反(fan)復(fu)凍(dong)融(rong)。?3.細(xi)(xi)胞培養(yang)物(wu)(wu)上清(qing)(qing)或(huo)(huo)(huo)(huo)(huo)(huo)(huo)(huo)其它生(sheng)物(wu)(wu)標(biao)(biao)(biao)(biao)本(ben)(ben)(ben)(ben)：1000 x g離(li)(li)心20分(fen)(fen)鐘(zhong)(zhong)，取(qu)(qu)上清(qing)(qing)即(ji)可檢(jian)(jian)測(ce)(ce)(ce)，或(huo)(huo)(huo)(huo)(huo)(huo)(huo)(huo)將(jiang)(jiang)標(biao)(biao)(biao)(biao)本(ben)(ben)(ben)(ben)放于(yu)(yu)-20℃或(huo)(huo)(huo)(huo)(huo)(huo)(huo)(huo)-80℃保(bao)(bao)存(cun)，但(dan)應避(bi)(bi)免(mian)(mian)反(fan)復(fu)凍(dong)融(rong)。 (注：標(biao)(biao)(biao)(biao)本(ben)(ben)(ben)(ben)溶(rong)血(xue)會影響**檢(jian)(jian)測(ce)(ce)(ce)結(jie)(jie)果，因(yin)(yin)此(ci)溶(rong)血(xue)標(biao)(biao)(biao)(biao)本(ben)(ben)(ben)(ben)不(bu)(bu)(bu)宜(yi)進行此(ci)項檢(jian)(jian)測(ce)(ce)(ce)。)???結(jie)(jie)果分(fen)(fen)析根(gen)據試(shi)(shi)(shi)劑(ji)盒提供的(de)(de)(de)(de)(de)(de)標(biao)(biao)(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)(pin)濃(nong)度(du)(du)(du)及(ji)對應的(de)(de)(de)(de)(de)(de)OD值，利用(yong)(yong)標(biao)(biao)(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)(pin)繪制(zhi)的(de)(de)(de)(de)(de)(de)標(biao)(biao)(biao)(biao)準(zhun)(zhun)曲(qu)線，計(ji)(ji)算(suan)待(dai)測(ce)(ce)(ce)樣(yang)(yang)品(pin)(pin)(pin)(pin)的(de)(de)(de)(de)(de)(de)濃(nong)度(du)(du)(du)。一(yi)般(ban)采(cai)(cai)用(yong)(yong)擬(ni)合(he)曲(qu)線法(fa)(fa)或(huo)(huo)(huo)(huo)(huo)(huo)(huo)(huo)直線回歸(gui)法(fa)(fa)進行數據處(chu)理，通過計(ji)(ji)算(suan)得到待(dai)測(ce)(ce)(ce)樣(yang)(yang)品(pin)(pin)(pin)(pin)的(de)(de)(de)(de)(de)(de)濃(nong)度(du)(du)(du)。???免(mian)(mian)責聲明試(shi)(shi)(shi)劑(ji)盒僅供研究(jiu)使(shi)用(yong)(yong)，不(bu)(bu)(bu)得用(yong)(yong)于(yu)(yu)臨床實(shi)驗(yan)(yan)(yan)(yan)(yan)(yan)或(huo)(huo)(huo)(huo)(huo)(huo)(huo)(huo)生(sheng)物(wu)(wu)體(ti)(ti)實(shi)驗(yan)(yan)(yan)(yan)(yan)(yan)，否則所(suo)產生(sheng)的(de)(de)(de)(de)(de)(de)一(yi)切(qie)后(hou)果，由(you)(you)實(shi)驗(yan)(yan)(yan)(yan)(yan)(yan)者承擔(dan)， 本(ben)(ben)(ben)(ben)公(gong)司概不(bu)(bu)(bu)負(fu)責。嚴格(ge)按照說明書(shu)操(cao)(cao)(cao)作(zuo)(zuo)(zuo)(zuo)，不(bu)(bu)(bu)同批號不(bu)(bu)(bu)可交(jiao)換使(shi)用(yong)(yong)，實(shi)驗(yan)(yan)(yan)(yan)(yan)(yan)者違反(fan)說明書(shu)操(cao)(cao)(cao)作(zuo)(zuo)(zuo)(zuo)，后(hou)果由(you)(you)實(shi)驗(yan)(yan)(yan)(yan)(yan)(yan)者承擔(dan)。?
?人(ren)(ren)P物(wu)(wu)(wu)(wu)質(SP)ELISA試(shi)(shi)(shi)(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)實(shi)(shi)(shi)驗(yan)原理? ? ? 試(shi)(shi)(shi)(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)采用(yong)(yong)雙抗(kang)體(ti)(ti)(ti)(ti)一(yi)(yi)步(bu)夾心(xin)法酶(mei)(mei)聯(lian)(lian)(lian)吸附試(shi)(shi)(shi)(shi)(shi)(shi)(shi)驗(yan)（ELISA）。往預先包被相(xiang)(xiang)關(guan)指標(biao)(biao)(biao)(biao)(biao)的(de)(de)(de)抗(kang)體(ti)(ti)(ti)(ti)的(de)(de)(de)包被微(wei)(wei)孔(kong)中，依次加入(ru)(ru)(ru)標(biao)(biao)(biao)(biao)(biao)本(ben)(ben)、標(biao)(biao)(biao)(biao)(biao)準品(pin)(pin)、HRP標(biao)(biao)(biao)(biao)(biao)記的(de)(de)(de)檢(jian)(jian)測(ce)(ce)抗(kang)體(ti)(ti)(ti)(ti)，經過溫育并洗(xi)滌。用(yong)(yong)底物(wu)(wu)(wu)(wu)TMB顯色，TMB在過氧(yang)化(hua)(hua)物(wu)(wu)(wu)(wu)酶(mei)(mei)的(de)(de)(de)催化(hua)(hua)下轉(zhuan)化(hua)(hua)成(cheng)藍色，并在酸的(de)(de)(de)作(zuo)(zuo)用(yong)(yong)下轉(zhuan)化(hua)(hua)成(cheng)終(zhong)的(de)(de)(de)黃色。顏色的(de)(de)(de)深淺和(he)樣(yang)品(pin)(pin)中的(de)(de)(de)相(xiang)(xiang)關(guan)指標(biao)(biao)(biao)(biao)(biao)呈正相(xiang)(xiang)關(guan)。用(yong)(yong)酶(mei)(mei)標(biao)(biao)(biao)(biao)(biao)儀(yi)在450nm 波(bo)(bo)長(chang)下測(ce)(ce)定吸光度（OD 值(zhi)），計算樣(yang)品(pin)(pin)濃度。試(shi)(shi)(shi)(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)性(xing)能1. 特異(yi)性(xing)高：由于(yu)(yu)是(shi)抗(kang)原抗(kang)體(ti)(ti)(ti)(ti)的(de)(de)(de)特異(yi)性(xing)結合(he)，因(yin)此試(shi)(shi)(shi)(shi)(shi)(shi)(shi)驗(yan)的(de)(de)(de)特異(yi)性(xing)很高，能夠排除其(qi)他(ta)物(wu)(wu)(wu)(wu)質的(de)(de)(de)干擾(rao)。2. 靈(ling)敏(min)(min)度高：由于(yu)(yu)酶(mei)(mei)的(de)(de)(de)放(fang)(fang)大(da)作(zuo)(zuo)用(yong)(yong)，使(shi)得試(shi)(shi)(shi)(shi)(shi)(shi)(shi)驗(yan)的(de)(de)(de)靈(ling)敏(min)(min)度大(da)大(da)提高，能夠檢(jian)(jian)測(ce)(ce)出(chu)微(wei)(wei)量的(de)(de)(de)抗(kang)原或(huo)(huo)(huo)抗(kang)體(ti)(ti)(ti)(ti)。3. 操(cao)(cao)作(zuo)(zuo)簡便：ELISA試(shi)(shi)(shi)(shi)(shi)(shi)(shi)驗(yan)操(cao)(cao)作(zuo)(zuo)相(xiang)(xiang)對簡單，容易(yi)掌握，適合(he)于(yu)(yu)大(da)規模樣(yang)本(ben)(ben)檢(jian)(jian)測(ce)(ce)。4. 適用(yong)(yong)范圍(wei)廣：可(ke)以用(yong)(yong)于(yu)(yu)檢(jian)(jian)測(ce)(ce)各種抗(kang)原、抗(kang)體(ti)(ti)(ti)(ti)和(he)激(ji)(ji)素等(deng)生(sheng)(sheng)物(wu)(wu)(wu)(wu)分子(zi)(zi)。試(shi)(shi)(shi)(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)組成(cheng)試(shi)(shi)(shi)(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)組成(cheng)48孔(kong)配(pei)置(zhi)96孔(kong)配(pei)置(zhi)保(bao)存(cun)(cun)(cun)說明(ming)書(shu)1份1份封板(ban)膜(mo)2片（48）2片（96）密封袋1個(ge)1個(ge)酶(mei)(mei)標(biao)(biao)(biao)(biao)(biao)包被板(ban)1×481×962-8℃保(bao)存(cun)(cun)(cun)標(biao)(biao)(biao)(biao)(biao)準品(pin)(pin)0.5ml×1瓶(ping)(ping)(ping)0.5ml×1瓶(ping)(ping)(ping)2-8℃保(bao)存(cun)(cun)(cun)標(biao)(biao)(biao)(biao)(biao)準品(pin)(pin)稀(xi)釋液(ye)(ye)(ye)(ye)1.5ml×1瓶(ping)(ping)(ping)1.5ml×1瓶(ping)(ping)(ping)2-8℃保(bao)存(cun)(cun)(cun)酶(mei)(mei)標(biao)(biao)(biao)(biao)(biao)試(shi)(shi)(shi)(shi)(shi)(shi)(shi)劑(ji)(ji)3 ml×1瓶(ping)(ping)(ping)6 ml×1瓶(ping)(ping)(ping)2-8℃保(bao)存(cun)(cun)(cun)樣(yang)品(pin)(pin)稀(xi)釋液(ye)(ye)(ye)(ye)3 ml×1瓶(ping)(ping)(ping)6 ml×1瓶(ping)(ping)(ping)2-8℃保(bao)存(cun)(cun)(cun)顯色劑(ji)(ji)A液(ye)(ye)(ye)(ye)3 ml×1瓶(ping)(ping)(ping)6 ml×1瓶(ping)(ping)(ping)2-8℃保(bao)存(cun)(cun)(cun)顯色劑(ji)(ji)B液(ye)(ye)(ye)(ye)3 ml×1瓶(ping)(ping)(ping)6 ml×1瓶(ping)(ping)(ping)2-8℃保(bao)存(cun)(cun)(cun)終(zhong)止(zhi)液(ye)(ye)(ye)(ye)3ml×1瓶(ping)(ping)(ping)6ml×1瓶(ping)(ping)(ping)2-8℃保(bao)存(cun)(cun)(cun)20倍濃縮洗(xi)滌液(ye)(ye)(ye)(ye)20ml×1瓶(ping)(ping)(ping)23ml×1瓶(ping)(ping)(ping)2-8℃保(bao)存(cun)(cun)(cun)操(cao)(cao)作(zuo)(zuo)步(bu)驟人(ren)(ren)P物(wu)(wu)(wu)(wu)質(SP)ELISA試(shi)(shi)(shi)(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)1. 對應(ying)板(ban)孔(kong)中加入(ru)(ru)(ru)100μL標(biao)(biao)(biao)(biao)(biao)準品(pin)(pin)工作(zuo)(zuo)液(ye)(ye)(ye)(ye)或(huo)(huo)(huo)樣(yang)本(ben)(ben)，37℃孵(fu)(fu)育90分鐘2. 棄掉(diao)板(ban)內(nei)(nei)液(ye)(ye)(ye)(ye)體(ti)(ti)(ti)(ti)后(hou)(hou)(hou)(hou)，立即加入(ru)(ru)(ru)100μL生(sheng)(sheng)物(wu)(wu)(wu)(wu)su化(hua)(hua)抗(kang)體(ti)(ti)(ti)(ti)工作(zuo)(zuo)液(ye)(ye)(ye)(ye)，37℃孵(fu)(fu)育60分鐘3. 棄掉(diao)板(ban)內(nei)(nei)液(ye)(ye)(ye)(ye)體(ti)(ti)(ti)(ti)，洗(xi)板(ban)3次4. 每孔(kong)加入(ru)(ru)(ru)100μL HRP酶(mei)(mei)結合(he)物(wu)(wu)(wu)(wu)工作(zuo)(zuo)液(ye)(ye)(ye)(ye)，37℃孵(fu)(fu)育30分鐘，棄掉(diao)板(ban)內(nei)(nei)液(ye)(ye)(ye)(ye)體(ti)(ti)(ti)(ti)，洗(xi)板(ban)5次5. 每孔(kong)加入(ru)(ru)(ru)90μL底物(wu)(wu)(wu)(wu)溶(rong)(rong)(rong)液(ye)(ye)(ye)(ye)，37℃孵(fu)(fu)育15分鐘左右6. 每孔(kong)加入(ru)(ru)(ru)50μL終(zhong)止(zhi)液(ye)(ye)(ye)(ye)7. 立即在450nm波(bo)(bo)長(chang)下讀數，處理數據(ju)標(biao)(biao)(biao)(biao)(biao)本(ben)(ben)的(de)(de)(de)采集及保(bao)存(cun)(cun)(cun)1.血(xue)清(qing)：全(quan)血(xue)標(biao)(biao)(biao)(biao)(biao)本(ben)(ben)請于(yu)(yu)室溫放(fang)(fang)置(zhi)2小時(shi)或(huo)(huo)(huo)4℃過一(yi)(yi)晚后(hou)(hou)(hou)(hou)于(yu)(yu)1000 x g離(li)心(xin)20分鐘，取上(shang)清(qing)即可(ke)檢(jian)(jian)測(ce)(ce)，或(huo)(huo)(huo)將(jiang)標(biao)(biao)(biao)(biao)(biao)本(ben)(ben)放(fang)(fang)于(yu)(yu)-20℃或(huo)(huo)(huo)-80℃保(bao)存(cun)(cun)(cun)，但應(ying)避免反(fan)復凍(dong)融。?2.血(xue)漿：可(ke)用(yong)(yong)EDTA或(huo)(huo)(huo)肝素作(zuo)(zuo)為(wei)抗(kang)凝劑(ji)(ji)，標(biao)(biao)(biao)(biao)(biao)本(ben)(ben)采集后(hou)(hou)(hou)(hou)30分鐘內(nei)(nei)于(yu)(yu)2 - 8° C 1000 x g離(li)心(xin)15分鐘，或(huo)(huo)(huo)將(jiang)標(biao)(biao)(biao)(biao)(biao)本(ben)(ben)放(fang)(fang)于(yu)(yu)-20℃或(huo)(huo)(huo)-80℃保(bao)存(cun)(cun)(cun)，但應(ying)避免反(fan)復凍(dong)融。?3.細(xi)(xi)胞(bao)培養物(wu)(wu)(wu)(wu)上(shang)清(qing)或(huo)(huo)(huo)其(qi)它生(sheng)(sheng)物(wu)(wu)(wu)(wu)標(biao)(biao)(biao)(biao)(biao)本(ben)(ben)：1000 x g離(li)心(xin)20分鐘，取上(shang)清(qing)即可(ke)檢(jian)(jian)測(ce)(ce)，或(huo)(huo)(huo)將(jiang)標(biao)(biao)(biao)(biao)(biao)本(ben)(ben)放(fang)(fang)于(yu)(yu)-20℃或(huo)(huo)(huo)-80℃保(bao)存(cun)(cun)(cun)，但應(ying)避免反(fan)復凍(dong)融。 (注：標(biao)(biao)(biao)(biao)(biao)本(ben)(ben)溶(rong)(rong)(rong)血(xue)會影響(xiang)**檢(jian)(jian)測(ce)(ce)結果，因(yin)此溶(rong)(rong)(rong)血(xue)標(biao)(biao)(biao)(biao)(biao)本(ben)(ben)不(bu)(bu)宜(yi)進行此項檢(jian)(jian)測(ce)(ce)。)?免責聲明(ming)試(shi)(shi)(shi)(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)僅(jin)供研究使(shi)用(yong)(yong)，不(bu)(bu)得用(yong)(yong)于(yu)(yu)臨床實(shi)(shi)(shi)驗(yan)或(huo)(huo)(huo)生(sheng)(sheng)物(wu)(wu)(wu)(wu)體(ti)(ti)(ti)(ti)實(shi)(shi)(shi)驗(yan)，否(fou)則所產生(sheng)(sheng)的(de)(de)(de)一(yi)(yi)切后(hou)(hou)(hou)(hou)果，由實(shi)(shi)(shi)驗(yan)者(zhe)承擔(dan)， 本(ben)(ben)公司概不(bu)(bu)負責。嚴(yan)格按照說明(ming)書(shu)操(cao)(cao)作(zuo)(zuo)，不(bu)(bu)同(tong)批號不(bu)(bu)可(ke)交換使(shi)用(yong)(yong)，實(shi)(shi)(shi)驗(yan)者(zhe)違反(fan)說明(ming)書(shu)操(cao)(cao)作(zuo)(zuo)，后(hou)(hou)(hou)(hou)果由實(shi)(shi)(shi)驗(yan)者(zhe)承擔(dan)。相(xiang)(xiang)關(guan)產品(pin)(pin)DM5394人(ren)(ren)微(wei)(wei)管關(guan)聯(lian)(lian)(lian)蛋(dan)(dan)白(MAPτ)酶(mei)(mei)聯(lian)(lian)(lian)免疫(yi)(yi)試(shi)(shi)(shi)(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)DM5395人(ren)(ren)TGF-β誘導(dao)早期應(ying)答基因(yin)1(TIEG1)酶(mei)(mei)聯(lian)(lian)(lian)免疫(yi)(yi)試(shi)(shi)(shi)(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)?DM5396人(ren)(ren)Toll樣(yang)受(shou)(shou)體(ti)(ti)(ti)(ti)2(TLR2)酶(mei)(mei)聯(lian)(lian)(lian)免疫(yi)(yi)試(shi)(shi)(shi)(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)DM5397人(ren)(ren)Toll樣(yang)受(shou)(shou)體(ti)(ti)(ti)(ti)9(TLR9)酶(mei)(mei)聯(lian)(lian)(lian)免疫(yi)(yi)試(shi)(shi)(shi)(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)DM5398人(ren)(ren)TU3A蛋(dan)(dan)白(TU3A)酶(mei)(mei)聯(lian)(lian)(lian)免疫(yi)(yi)試(shi)(shi)(shi)(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)DM5399人(ren)(ren)蛋(dan)(dan)白酶(mei)(mei)激(ji)(ji)活受(shou)(shou)體(ti)(ti)(ti)(ti)3(PAR3)酶(mei)(mei)聯(lian)(lian)(lian)免疫(yi)(yi)試(shi)(shi)(shi)(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)DM5400人(ren)(ren)T細(xi)(xi)胞(bao)活化(hua)(hua)連接(jie)蛋(dan)(dan)白(LAT)酶(mei)(mei)聯(lian)(lian)(lian)免疫(yi)(yi)試(shi)(shi)(shi)(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)DM5401人(ren)(ren)T細(xi)(xi)胞(bao)急性(xing)淋巴(ba)母細(xi)(xi)胞(bao)白血(xue)病(bing)相(xiang)(xiang)關(guan)抗(kang)原(TALLA-1/CD231)酶(mei)(mei)聯(lian)(lian)(lian)免疫(yi)(yi)試(shi)(shi)(shi)(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)DM5402人(ren)(ren)T細(xi)(xi)胞(bao)生(sheng)(sheng)長(chang)因(yin)子(zi)(zi)-Ⅲ(TCGF-Ⅲ)酶(mei)(mei)聯(lian)(lian)(lian)免疫(yi)(yi)試(shi)(shi)(shi)(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)?DM5403人(ren)(ren)T細(xi)(xi)胞(bao)受(shou)(shou)體(ti)(ti)(ti)(ti)(TCR)酶(mei)(mei)聯(lian)(lian)(lian)免疫(yi)(yi)試(shi)(shi)(shi)(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)DM5404人(ren)(ren)T細(xi)(xi)胞(bao)特異(yi)性(xing)鳥苷三磷酸酶(mei)(mei)(Tgtp)酶(mei)(mei)聯(lian)(lian)(lian)免疫(yi)(yi)試(shi)(shi)(shi)(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)DM5405人(ren)(ren)β-骨膠原交聯(lian)(lian)(lian)(β-CTx)酶(mei)(mei)聯(lian)(lian)(lian)免疫(yi)(yi)試(shi)(shi)(shi)(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)
本(ben)(ben)(ben)(ben)試(shi)(shi)(shi)劑(ji)盒(he)只能(neng)用(yong)于(yu)(yu)科學研究(jiu)，不得(de)用(yong)于(yu)(yu)醫學診斷人（Human）甲(jia)胎(tai)蛋(dan)白(bai)(bai)（AFP）ELISA檢(jian)(jian)(jian)測(ce)(ce)試(shi)(shi)(shi)劑(ji)盒(he)使(shi)用(yong)說(shuo)(shuo)明書(shu)檢(jian)(jian)(jian)測(ce)(ce)原(yuan)理(li)試(shi)(shi)(shi)劑(ji)盒(he)采用(yong)雙抗(kang)體(ti)(ti)一(yi)步(bu)夾心(xin)法酶聯免疫吸(xi)(xi)(xi)附(fu)試(shi)(shi)(shi)驗(yan)（ELISA）。往(wang)預先包被甲(jia)胎(tai)蛋(dan)白(bai)(bai)（AFP）抗(kang)體(ti)(ti)的(de)(de)(de)(de)(de)包被微(wei)(wei)孔(kong)(kong)(kong)(kong)(kong)(kong)中(zhong)，依次(ci)加(jia)(jia)(jia)入標(biao)(biao)(biao)(biao)(biao)本(ben)(ben)(ben)(ben)、標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)、HRP標(biao)(biao)(biao)(biao)(biao)記的(de)(de)(de)(de)(de)檢(jian)(jian)(jian)測(ce)(ce)抗(kang)體(ti)(ti)，經過(guo)(guo)溫(wen)(wen)育并徹底(di)洗(xi)(xi)(xi)滌(di)。用(yong)底(di)物(wu)(wu)TMB顯(xian)色(se)，TMB在(zai)過(guo)(guo)氧化(hua)(hua)物(wu)(wu)酶的(de)(de)(de)(de)(de)催(cui)化(hua)(hua)下轉(zhuan)(zhuan)化(hua)(hua)成(cheng)藍色(se)，并在(zai)酸(suan)(suan)的(de)(de)(de)(de)(de)作(zuo)(zuo)用(yong)下轉(zhuan)(zhuan)化(hua)(hua)成(cheng)＊終(zhong)的(de)(de)(de)(de)(de)黃(huang)色(se)。顏色(se)的(de)(de)(de)(de)(de)深淺和(he)樣(yang)(yang)(yang)(yang)品(pin)(pin)(pin)(pin)(pin)中(zhong)的(de)(de)(de)(de)(de)甲(jia)胎(tai)蛋(dan)白(bai)(bai)（AFP）呈正相關(guan)。用(yong)酶標(biao)(biao)(biao)(biao)(biao)儀(yi)在(zai)450nm 波(bo)長(chang)下測(ce)(ce)定吸(xi)(xi)(xi)光(guang)度(du)（OD 值(zhi)），計(ji)算(suan)樣(yang)(yang)(yang)(yang)品(pin)(pin)(pin)(pin)(pin)濃度(du)。樣(yang)(yang)(yang)(yang)品(pin)(pin)(pin)(pin)(pin)收(shou)集、處理(li)及保(bao)存(cun)(cun)方(fang)法1. 血(xue)(xue)清(qing)：使(shi)用(yong)不含熱原(yuan)和(he)內(nei)(nei)毒素的(de)(de)(de)(de)(de)試(shi)(shi)(shi)管，操作(zuo)(zuo)過(guo)(guo)程中(zhong)避(bi)免任何細胞(bao)刺激，收(shou)集血(xue)(xue)液(ye)(ye)(ye)(ye)后(hou)，3000轉(zhuan)(zhuan)離心(xin)10分(fen)鐘(zhong)將血(xue)(xue)清(qing)和(he)紅(hong)細胞(bao)迅速小(xiao)(xiao)心(xin)地分(fen)離。2. 血(xue)(xue)漿：EDTA、檸(ning)檬酸(suan)(suan)鹽或肝素抗(kang)凝。3000轉(zhuan)(zhuan)離心(xin)30分(fen)鐘(zhong)取上清(qing)。3. 細胞(bao)上清(qing)液(ye)(ye)(ye)(ye)：3000轉(zhuan)(zhuan)離心(xin)10分(fen)鐘(zhong)去(qu)(qu)除顆粒和(he)聚合物(wu)(wu)。4. 組(zu)(zu)(zu)織(zhi)勻漿：將組(zu)(zu)(zu)織(zhi)加(jia)(jia)(jia)入適(shi)量生(sheng)(sheng)理(li)鹽水(shui)(shui)搗碎。3000轉(zhuan)(zhuan)離心(xin)10分(fen)鐘(zhong)取上清(qing)。5. 保(bao)存(cun)(cun)：如(ru)果樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)收(shou)集后(hou)不及時檢(jian)(jian)(jian)測(ce)(ce)，請按(an)(an)一(yi)次(ci)用(yong)量分(fen)裝，凍(dong)存(cun)(cun)于(yu)(yu)-20℃，避(bi)免反(fan)復凍(dong)融，在(zai)室溫(wen)(wen)下解凍(dong)并確(que)保(bao)樣(yang)(yang)(yang)(yang)品(pin)(pin)(pin)(pin)(pin)均(jun)勻地充(chong)分(fen)解凍(dong)。自(zi)備物(wu)(wu)品(pin)(pin)(pin)(pin)(pin)1. 酶標(biao)(biao)(biao)(biao)(biao)儀(yi)（450nm）2. 高精度(du)加(jia)(jia)(jia)樣(yang)(yang)(yang)(yang)器(qi)及槍(qiang)頭(tou)：0.5-10uL、2-20uL、20-200uL、200-1000uL3. 37℃恒溫(wen)(wen)箱操作(zuo)(zuo)注(zhu)意事項1. 試(shi)(shi)(shi)劑(ji)盒(he)保(bao)存(cun)(cun)在(zai)2-8℃，使(shi)用(yong)前(qian)室溫(wen)(wen)平衡20分(fen)鐘(zhong)。從冰箱取出(chu)的(de)(de)(de)(de)(de)濃縮洗(xi)(xi)(xi)滌(di)液(ye)(ye)(ye)(ye)會有結(jie)晶(jing)，這(zhe)屬于(yu)(yu)正常(chang)現象，水(shui)(shui)浴(yu)加(jia)(jia)(jia)熱使(shi)結(jie)晶(jing)完全溶解后(hou)再(zai)使(shi)用(yong)。2. 實(shi)驗(yan)中(zhong)不用(yong)的(de)(de)(de)(de)(de)板(ban)(ban)(ban)條(tiao)應(ying)(ying)立即放回(hui)自(zi)封袋(dai)中(zhong)，密封（低溫(wen)(wen)干(gan)燥）保(bao)存(cun)(cun)。3. 濃度(du)為0的(de)(de)(de)(de)(de)S0號標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)即可視為陰(yin)性(xing)對照(zhao)(zhao)或者(zhe)空(kong)(kong)白(bai)(bai)；按(an)(an)照(zhao)(zhao)說(shuo)(shuo)明書(shu)操作(zuo)(zuo)時樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)已經稀釋(shi)5倍，＊終(zhong)結(jie)果乘以5才是樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)實(shi)際濃度(du)。4. 嚴格(ge)按(an)(an)照(zhao)(zhao)說(shuo)(shuo)明書(shu)中(zhong)標(biao)(biao)(biao)(biao)(biao)明的(de)(de)(de)(de)(de)時間(jian)、加(jia)(jia)(jia)液(ye)(ye)(ye)(ye)量及順序進行溫(wen)(wen)育操作(zuo)(zuo)。5. 所有液(ye)(ye)(ye)(ye)體(ti)(ti)組(zu)(zu)(zu)分(fen)使(shi)用(yong)前(qian)充(chong)分(fen)搖勻。試(shi)(shi)(shi)劑(ji)盒(he)組(zu)(zu)(zu)成(cheng)名稱96孔(kong)(kong)(kong)(kong)(kong)(kong)配(pei)置48孔(kong)(kong)(kong)(kong)(kong)(kong)配(pei)置備注(zhu)微(wei)(wei)孔(kong)(kong)(kong)(kong)(kong)(kong)酶標(biao)(biao)(biao)(biao)(biao)板(ban)(ban)(ban)12孔(kong)(kong)(kong)(kong)(kong)(kong)×8條(tiao)12孔(kong)(kong)(kong)(kong)(kong)(kong)×4條(tiao)無(wu)(wu)(wu)(wu)標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)0.3mL*6管0.3mL*6管無(wu)(wu)(wu)(wu)樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)稀釋(shi)液(ye)(ye)(ye)(ye)6mL3mL無(wu)(wu)(wu)(wu)檢(jian)(jian)(jian)測(ce)(ce)抗(kang)體(ti)(ti)-HRP10mL5mL無(wu)(wu)(wu)(wu)20×洗(xi)(xi)(xi)滌(di)緩(huan)沖(chong)液(ye)(ye)(ye)(ye)25mL15mL按(an)(an)說(shuo)(shuo)明書(shu)進行稀釋(shi)底(di)物(wu)(wu)A6mL3mL無(wu)(wu)(wu)(wu)底(di)物(wu)(wu)B6mL3mL無(wu)(wu)(wu)(wu)終(zhong)止液(ye)(ye)(ye)(ye)6mL3mL無(wu)(wu)(wu)(wu)封板(ban)(ban)(ban)膜2張2張無(wu)(wu)(wu)(wu)說(shuo)(shuo)明書(shu)1份(fen)1份(fen)無(wu)(wu)(wu)(wu)自(zi)封袋(dai)1個(ge)1個(ge)無(wu)(wu)(wu)(wu)注(zhu)：標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)（S0-S5）濃度(du)依次(ci)為：0、2、4、8、16、32 ng/mL試(shi)(shi)(shi)劑(ji)的(de)(de)(de)(de)(de)準(zhun)(zhun)(zhun)(zhun)(zhun)備 20×洗(xi)(xi)(xi)滌(di)緩(huan)沖(chong)液(ye)(ye)(ye)(ye)的(de)(de)(de)(de)(de)稀釋(shi)：蒸(zheng)餾水(shui)(shui)按(an)(an)1：20稀釋(shi)，即1份(fen)的(de)(de)(de)(de)(de)20×洗(xi)(xi)(xi)滌(di)緩(huan)沖(chong)液(ye)(ye)(ye)(ye)加(jia)(jia)(jia)19份(fen)的(de)(de)(de)(de)(de)蒸(zheng)餾水(shui)(shui)。洗(xi)(xi)(xi)板(ban)(ban)(ban)方(fang)法1. 手工洗(xi)(xi)(xi)板(ban)(ban)(ban)：甩盡孔(kong)(kong)(kong)(kong)(kong)(kong)內(nei)(nei)液(ye)(ye)(ye)(ye)體(ti)(ti)，每(mei)孔(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)(jia)滿洗(xi)(xi)(xi)滌(di)液(ye)(ye)(ye)(ye)，靜置1min后(hou)甩盡孔(kong)(kong)(kong)(kong)(kong)(kong)內(nei)(nei)液(ye)(ye)(ye)(ye)體(ti)(ti)，在(zai)吸(xi)(xi)(xi)水(shui)(shui)紙上拍干(gan)，如(ru)此洗(xi)(xi)(xi)板(ban)(ban)(ban)5次(ci)。2. 自(zi)動(dong)洗(xi)(xi)(xi)板(ban)(ban)(ban)機(ji)：每(mei)孔(kong)(kong)(kong)(kong)(kong)(kong)注(zhu)入洗(xi)(xi)(xi)液(ye)(ye)(ye)(ye)350μL，浸泡1min，洗(xi)(xi)(xi)板(ban)(ban)(ban)5次(ci)。操作(zuo)(zuo)步(bu)驟1. 從室溫(wen)(wen)平衡20min后(hou)的(de)(de)(de)(de)(de)鋁(lv)箔袋(dai)中(zhong)取出(chu)所需板(ban)(ban)(ban)條(tiao)，剩余板(ban)(ban)(ban)條(tiao)用(yong)自(zi)封袋(dai)密封放回(hui)4℃。2. 設置標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)孔(kong)(kong)(kong)(kong)(kong)(kong)和(he)樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)孔(kong)(kong)(kong)(kong)(kong)(kong)，標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)孔(kong)(kong)(kong)(kong)(kong)(kong)各加(jia)(jia)(jia)不同濃度(du)的(de)(de)(de)(de)(de)標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)50μL；3. 樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)孔(kong)(kong)(kong)(kong)(kong)(kong)先加(jia)(jia)(jia)待測(ce)(ce)樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)10μL，再(zai)加(jia)(jia)(jia)樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)稀釋(shi)液(ye)(ye)(ye)(ye)40μL；空(kong)(kong)白(bai)(bai)孔(kong)(kong)(kong)(kong)(kong)(kong)不加(jia)(jia)(jia)。4. 除空(kong)(kong)白(bai)(bai)孔(kong)(kong)(kong)(kong)(kong)(kong)外，標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)孔(kong)(kong)(kong)(kong)(kong)(kong)和(he)樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)孔(kong)(kong)(kong)(kong)(kong)(kong)中(zhong)每(mei)孔(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)(jia)入辣根過(guo)(guo)氧化(hua)(hua)物(wu)(wu)酶（HRP）標(biao)(biao)(biao)(biao)(biao)記的(de)(de)(de)(de)(de)檢(jian)(jian)(jian)測(ce)(ce)抗(kang)體(ti)(ti)100μL，用(yong)封板(ban)(ban)(ban)膜封住反(fan)應(ying)(ying)孔(kong)(kong)(kong)(kong)(kong)(kong)，37℃水(shui)(shui)浴(yu)鍋或恒溫(wen)(wen)箱溫(wen)(wen)育60min。5. 棄去(qu)(qu)液(ye)(ye)(ye)(ye)體(ti)(ti)，吸(xi)(xi)(xi)水(shui)(shui)紙上拍干(gan)，每(mei)孔(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)(jia)滿洗(xi)(xi)(xi)滌(di)液(ye)(ye)(ye)(ye)，靜置1min，甩去(qu)(qu)洗(xi)(xi)(xi)滌(di)液(ye)(ye)(ye)(ye)，吸(xi)(xi)(xi)水(shui)(shui)紙上拍干(gan)，如(ru)此重復洗(xi)(xi)(xi)板(ban)(ban)(ban)5次(ci)（也(ye)可用(yong)洗(xi)(xi)(xi)板(ban)(ban)(ban)機(ji)洗(xi)(xi)(xi)板(ban)(ban)(ban)）。6. 每(mei)孔(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)(jia)入底(di)物(wu)(wu)A、B各50μL，37℃避(bi)光(guang)孵育15min。7. 每(mei)孔(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)(jia)入終(zhong)止液(ye)(ye)(ye)(ye)50μL，15min內(nei)(nei)，在(zai)450nm波(bo)長(chang)處測(ce)(ce)定各孔(kong)(kong)(kong)(kong)(kong)(kong)的(de)(de)(de)(de)(de)OD值(zhi)。結(jie)果判斷 繪制標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)曲線：在(zai)Excel工作(zuo)(zuo)表中(zhong)，以標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)濃度(du)作(zuo)(zuo)橫(heng)坐標(biao)(biao)(biao)(biao)(biao)，對應(ying)(ying)OD值(zhi)作(zuo)(zuo)縱坐標(biao)(biao)(biao)(biao)(biao)，繪制出(chu)標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)線性(xing)回(hui)歸曲線，按(an)(an)曲線方(fang)程計(ji)算(suan)各樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)濃度(du)值(zhi)。 試(shi)(shi)(shi)劑(ji)盒(he)性(xing)能(neng)1. 準(zhun)(zhun)(zhun)(zhun)(zhun)確(que)性(xing)：標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)線性(xing)回(hui)歸與預期(qi)濃度(du)相關(guan)系數R值(zhi)，大于(yu)(yu)等于(yu)(yu)0.9900。2. 靈敏度(du)：＊低檢(jian)(jian)(jian)測(ce)(ce)濃度(du)小(xiao)(xiao)于(yu)(yu)0.1 ng/mL。3. 特(te)異(yi)性(xing)：不與其它可溶性(xing)結(jie)構(gou)類似物(wu)(wu)交叉反(fan)應(ying)(ying)。4. 重復性(xing)：板(ban)(ban)(ban)內(nei)(nei)、板(ban)(ban)(ban)間(jian)變異(yi)系數均(jun)小(xiao)(xiao)于(yu)(yu)15%。5. 貯藏：2-8℃，避(bi)光(guang)防(fang)潮保(bao)存(cun)(cun)。6. 有效期(qi)：6個(ge)月免責(ze)聲(sheng)明1. 試(shi)(shi)(shi)劑(ji)盒(he)僅供(gong)研究(jiu)使(shi)用(yong)，不得(de)用(yong)于(yu)(yu)臨床實(shi)驗(yan)或人體(ti)(ti)實(shi)驗(yan)，否則所產生(sheng)(sheng)的(de)(de)(de)(de)(de)一(yi)切后(hou)果，由(you)實(shi)驗(yan)者(zhe)承擔，本(ben)(ben)(ben)(ben)公司概不負(fu)責(ze)。2. 嚴格(ge)按(an)(an)照(zhao)(zhao)說(shuo)(shuo)明書(shu)操作(zuo)(zuo)，實(shi)驗(yan)者(zhe)違反(fan)說(shuo)(shuo)明書(shu)操作(zuo)(zuo)，后(hou)果由(you)實(shi)驗(yan)者(zhe)承擔。 FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES. Human Alpha-fetoprotein (AFP) ELISA Kit instruction Intended useThis AFP ELISA kit is intended Laboratory for Research use only and is not for use in diagnostic or therapeutic procedures.The Stop Solution changes the color from blue to yellow and the intensity of the color is measured at 450 nm using a spectrophotometer. In order to measure the concentration of AFP in the sample, this AFP ELISA Kit includes a set of calibration standards. The calibration standards are assayed at the same time as the samples and allow the operator to produce a standard curve of Optical Density versus AFP concentration. The concentration of AFP in the samples is then determined by comparing the O.D. of the samples to the standard curve.Sample collection and storagesSerum - Use a serum separator tube and allow samples to clot for 30 minutes before centrifugation for 10 minutes at approximately 3000×g. Remove serum and assay immediately or aliquot and store samples at -20℃ or -80℃.Avoid repeated freeze-thaw cyclesPlasma - Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 30 minutes at 3000×g at 2-8℃ within 30 minutes of collection. Store samples at -20℃or -80℃. Avoid repeated freeze-thaw cycles.Cell culture supernates and other biological fluids - Remove particulates by centrifugation and assay immediately or aliquot and store samples at -20℃or -80℃. Avoid repeated freeze-thaw cycles.Note: The samples shoule be centrifugated dequately and no hemolysis or granule was allowed.Materials required but not supplied1. Standard microplate reader(450nm)2. Precision pipettes and Disposable pipette tips.3. 37 ℃ incubatorPrecautions1. Do not substitute reagents from one kit to another. Standard, conjugate and microplates are matched for optimal performance. Use only the reagents supplied by manufacturer.2. Do not remove microplate from the storage bag until needed. Unused strips should be stored at 2-8°C in their pouch with the desiccant provided.3. Mix all reagents before using.Remove all kit reagents from refrigerator and allow them to reach room temperature ( 20-25°C)Materials suppliedName96 determinations48 determinationsMicroelisa stripplate12*8strips12*4stripsStandard0.3ml*6tubes0.3ml*6tubesSample Diluent6.0ml3.0mlHRP-Conjugate reagent10.0ml5.0ml20X Wash solution25ml15mlChromogen Solution A6.0ml3.0mlChromogen Solution B6.0ml3.0mlStop Solution6.0ml3.0mlClosure plate membrane22User manual11Sealed bags11Note: Standard (S0 → S5) concentration was followed by:0,2,4,8,16,32 ng/ml.Reagent preparation20×wash solution:Dilute with Distilled or deionized water 1:20.Assay procedure1. Prepare all reagents before starting assay procedure. It is recommended that all Standards and Samples be added in duplicate to the Microelisa Stripplate.2. Add standard: Set Standard wells, testing sample wells. Add standard 50μl to standard well.3. Add Sample: Add testing sample 10μl then add Sample Diluent 40μl to testing sample well; Blank well doesn’t add anyting.4. Add 100μl of HRP-conjugate reagent to each well, cover with an adhesive strip and incubate for 60 minutes at 37°C. 5. Aspirate each well and wash, repeating the process four times for a total of five washes. Wash by filling each well with Wash Solution (400μl) using a squirt bottle, manifold dispenser or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Solution by aspirating or decanting. Invert the plate and blot it against clean paper towels.6. Add chromogen solution A 50μl and chromogen solution B 50μl to each well. Gently mix and incubate for 15 minutes at 37°C. Protect from light.7. Add 50μl Stop Solution to each well. The color in the wells should change from blue to yellow. If the color in the wells is green or the color change does not appear uniform, gently tap the plate to ensure thorough mixing.8. Read the Optical Density (O.D.) at 450 nm using a microtiter plate reader within 15 minutes.Calculation of results1. This standard curve is used to determine the amount in an unknown sample. The standard curve is generated by plotting the average O.D. (450 nm) obtained for each of the six standard concentrations on the vertical (Y) axis versus the corresponding concentration on the horizontal (X) axis. 2. First, calculate the mean O.D. value for each standard and sample. All O.D. values, are subtracted by the mean value of the zero standard before result interpretation. Construct the standard curve using graph paper or statistical software. 3. To determine the amount in each sample, first locate the O.D. value on the Y-axis and extend a horizontal line to the standard curve. At the point of intersection, draw a vertical line to the X-axis and read the corresponding concentration. 4. Any variation in operator, pipetting and washing technique, incubation time or temperature, and kit age can cause variation in result. Each user should obtain their own standard curve.5. The sensitivity by this assay is 0.1 ng/ml6. Standard curve Storage： 2-8℃.validity： six months. FOR RESEARCH USE ONLY; NOT FOR THERAPEUTIC OR DIAGNOSTIC APPLICATIONS! PLEASE READ THROUGH ENTIRE PROCEDURE BEFORE BEGINNING!
本(ben)(ben)試(shi)(shi)(shi)劑(ji)(ji)(ji)盒(he)只(zhi)能用(yong)(yong)(yong)(yong)于(yu)科學研究，不(bu)得用(yong)(yong)(yong)(yong)于(yu)醫學診斷(duan)人（Human）白蛋(dan)白（ALB）ELISA檢(jian)(jian)測試(shi)(shi)(shi)劑(ji)(ji)(ji)盒(he)使用(yong)(yong)(yong)(yong)說(shuo)明書檢(jian)(jian)測原理試(shi)(shi)(shi)劑(ji)(ji)(ji)盒(he)采用(yong)(yong)(yong)(yong)雙抗(kang)體(ti)(ti)一步夾心(xin)(xin)(xin)法酶(mei)聯免疫吸附試(shi)(shi)(shi)驗(yan)(yan)（ELISA）。往(wang)預先(xian)包(bao)被白蛋(dan)白（ALB）抗(kang)體(ti)(ti)的(de)(de)(de)(de)包(bao)被微孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)中(zhong)，依次加(jia)入(ru)(ru)(ru)標(biao)(biao)(biao)本(ben)(ben)、標(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)、HRP標(biao)(biao)(biao)記的(de)(de)(de)(de)檢(jian)(jian)測抗(kang)體(ti)(ti)，經過(guo)(guo)溫育(yu)(yu)并徹底(di)洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)。用(yong)(yong)(yong)(yong)底(di)物(wu)(wu)TMB顯(xian)色(se)(se)，TMB在(zai)過(guo)(guo)氧化(hua)(hua)物(wu)(wu)酶(mei)的(de)(de)(de)(de)催化(hua)(hua)下(xia)轉(zhuan)(zhuan)化(hua)(hua)成(cheng)藍(lan)色(se)(se)，并在(zai)酸的(de)(de)(de)(de)作(zuo)用(yong)(yong)(yong)(yong)下(xia)轉(zhuan)(zhuan)化(hua)(hua)成(cheng)＊終(zhong)(zhong)的(de)(de)(de)(de)黃色(se)(se)。顏色(se)(se)的(de)(de)(de)(de)深淺和(he)(he)樣(yang)品(pin)(pin)(pin)中(zhong)的(de)(de)(de)(de)白蛋(dan)白（ALB）呈正(zheng)相關。用(yong)(yong)(yong)(yong)酶(mei)標(biao)(biao)(biao)儀在(zai)450nm 波長(chang)下(xia)測定吸光度(du)（OD 值(zhi)），計算樣(yang)品(pin)(pin)(pin)濃(nong)(nong)度(du)。樣(yang)品(pin)(pin)(pin)收集(ji)、處理及保(bao)存(cun)方法1. 血清(qing)(qing)：使用(yong)(yong)(yong)(yong)不(bu)含熱原和(he)(he)內毒素的(de)(de)(de)(de)試(shi)(shi)(shi)管(guan)(guan)，操作(zuo)過(guo)(guo)程中(zhong)避免任何細胞(bao)刺激，收集(ji)血液(ye)(ye)(ye)(ye)后(hou)，3000轉(zhuan)(zhuan)離(li)(li)(li)心(xin)(xin)(xin)10分(fen)(fen)(fen)(fen)鐘(zhong)將血清(qing)(qing)和(he)(he)紅細胞(bao)迅速小心(xin)(xin)(xin)地分(fen)(fen)(fen)(fen)離(li)(li)(li)。2. 血漿(jiang)：EDTA、檸(ning)檬酸鹽(yan)或肝素抗(kang)凝。3000轉(zhuan)(zhuan)離(li)(li)(li)心(xin)(xin)(xin)30分(fen)(fen)(fen)(fen)鐘(zhong)取上(shang)(shang)清(qing)(qing)。3. 細胞(bao)上(shang)(shang)清(qing)(qing)液(ye)(ye)(ye)(ye)：3000轉(zhuan)(zhuan)離(li)(li)(li)心(xin)(xin)(xin)10分(fen)(fen)(fen)(fen)鐘(zhong)去除顆(ke)粒和(he)(he)聚合物(wu)(wu)。4. 組(zu)織(zhi)勻漿(jiang)：將組(zu)織(zhi)加(jia)入(ru)(ru)(ru)適量(liang)生理鹽(yan)水(shui)(shui)(shui)搗碎。3000轉(zhuan)(zhuan)離(li)(li)(li)心(xin)(xin)(xin)10分(fen)(fen)(fen)(fen)鐘(zhong)取上(shang)(shang)清(qing)(qing)。5. 保(bao)存(cun)：如果(guo)(guo)樣(yang)本(ben)(ben)收集(ji)后(hou)不(bu)及時(shi)檢(jian)(jian)測，請按一次用(yong)(yong)(yong)(yong)量(liang)分(fen)(fen)(fen)(fen)裝，凍(dong)(dong)存(cun)于(yu)-20℃，避免反復(fu)凍(dong)(dong)融，在(zai)室(shi)溫下(xia)解凍(dong)(dong)并確保(bao)樣(yang)品(pin)(pin)(pin)均(jun)勻地充分(fen)(fen)(fen)(fen)解凍(dong)(dong)。自備物(wu)(wu)品(pin)(pin)(pin)1. 酶(mei)標(biao)(biao)(biao)儀（450nm）2. 高精度(du)加(jia)樣(yang)器及槍頭(tou)：0.5-10uL、2-20uL、20-200uL、200-1000uL3. 37℃恒(heng)溫箱操作(zuo)注意(yi)事項1. 試(shi)(shi)(shi)劑(ji)(ji)(ji)盒(he)保(bao)存(cun)在(zai)2-8℃，使用(yong)(yong)(yong)(yong)前(qian)(qian)室(shi)溫平(ping)衡20分(fen)(fen)(fen)(fen)鐘(zhong)。從冰箱取出的(de)(de)(de)(de)濃(nong)(nong)縮洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)液(ye)(ye)(ye)(ye)會有結(jie)晶(jing)，這屬(shu)于(yu)正(zheng)常現象，水(shui)(shui)(shui)浴(yu)加(jia)熱使結(jie)晶(jing)完全溶(rong)解后(hou)再使用(yong)(yong)(yong)(yong)。2. 實(shi)(shi)(shi)驗(yan)(yan)中(zhong)不(bu)用(yong)(yong)(yong)(yong)的(de)(de)(de)(de)板(ban)(ban)(ban)(ban)條(tiao)應(ying)立即(ji)放(fang)回(hui)自封(feng)(feng)(feng)(feng)袋(dai)(dai)中(zhong)，密封(feng)(feng)(feng)(feng)（低溫干燥）保(bao)存(cun)。3. 濃(nong)(nong)度(du)為(wei)(wei)0的(de)(de)(de)(de)S0號(hao)標(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)即(ji)可(ke)(ke)視為(wei)(wei)陰性(xing)(xing)對照(zhao)或者空(kong)白；按照(zhao)說(shuo)明書操作(zuo)時(shi)樣(yang)本(ben)(ben)已經稀(xi)(xi)釋(shi)(shi)5倍，＊終(zhong)(zhong)結(jie)果(guo)(guo)乘以5才是(shi)樣(yang)本(ben)(ben)實(shi)(shi)(shi)際濃(nong)(nong)度(du)。4. 嚴格(ge)按照(zhao)說(shuo)明書中(zhong)標(biao)(biao)(biao)明的(de)(de)(de)(de)時(shi)間、加(jia)液(ye)(ye)(ye)(ye)量(liang)及順序進行(xing)溫育(yu)(yu)操作(zuo)。5. 所(suo)有液(ye)(ye)(ye)(ye)體(ti)(ti)組(zu)分(fen)(fen)(fen)(fen)使用(yong)(yong)(yong)(yong)前(qian)(qian)充分(fen)(fen)(fen)(fen)搖(yao)勻。試(shi)(shi)(shi)劑(ji)(ji)(ji)盒(he)組(zu)成(cheng)名稱96孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)配(pei)置(zhi)48孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)配(pei)置(zhi)備注微孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)酶(mei)標(biao)(biao)(biao)板(ban)(ban)(ban)(ban)12孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)×8條(tiao)12孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)×4條(tiao)無(wu)(wu)標(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)0.3mL*6管(guan)(guan)0.3mL*6管(guan)(guan)無(wu)(wu)樣(yang)本(ben)(ben)稀(xi)(xi)釋(shi)(shi)液(ye)(ye)(ye)(ye)6mL3mL無(wu)(wu)檢(jian)(jian)測抗(kang)體(ti)(ti)-HRP10mL5mL無(wu)(wu)20×洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)緩(huan)沖(chong)液(ye)(ye)(ye)(ye)25mL15mL按說(shuo)明書進行(xing)稀(xi)(xi)釋(shi)(shi)底(di)物(wu)(wu)A6mL3mL無(wu)(wu)底(di)物(wu)(wu)B6mL3mL無(wu)(wu)終(zhong)(zhong)止液(ye)(ye)(ye)(ye)6mL3mL無(wu)(wu)封(feng)(feng)(feng)(feng)板(ban)(ban)(ban)(ban)膜2張2張無(wu)(wu)說(shuo)明書1份1份無(wu)(wu)自封(feng)(feng)(feng)(feng)袋(dai)(dai)1個1個無(wu)(wu)注：標(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)（S0-S5）濃(nong)(nong)度(du)依次為(wei)(wei)：0、6.25、12.5、25、50、100 mg/mL試(shi)(shi)(shi)劑(ji)(ji)(ji)的(de)(de)(de)(de)準(zhun)(zhun)(zhun)(zhun)(zhun)備 20×洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)緩(huan)沖(chong)液(ye)(ye)(ye)(ye)的(de)(de)(de)(de)稀(xi)(xi)釋(shi)(shi)：蒸餾水(shui)(shui)(shui)按1：20稀(xi)(xi)釋(shi)(shi)，即(ji)1份的(de)(de)(de)(de)20×洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)緩(huan)沖(chong)液(ye)(ye)(ye)(ye)加(jia)19份的(de)(de)(de)(de)蒸餾水(shui)(shui)(shui)。洗(xi)(xi)(xi)(xi)(xi)板(ban)(ban)(ban)(ban)方法1. 手工洗(xi)(xi)(xi)(xi)(xi)板(ban)(ban)(ban)(ban)：甩盡孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)內液(ye)(ye)(ye)(ye)體(ti)(ti)，每孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)加(jia)滿洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)液(ye)(ye)(ye)(ye)，靜置(zhi)1min后(hou)甩盡孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)內液(ye)(ye)(ye)(ye)體(ti)(ti)，在(zai)吸水(shui)(shui)(shui)紙(zhi)(zhi)上(shang)(shang)拍干，如此洗(xi)(xi)(xi)(xi)(xi)板(ban)(ban)(ban)(ban)5次。2. 自動洗(xi)(xi)(xi)(xi)(xi)板(ban)(ban)(ban)(ban)機(ji)：每孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)注入(ru)(ru)(ru)洗(xi)(xi)(xi)(xi)(xi)液(ye)(ye)(ye)(ye)350μL，浸泡1min，洗(xi)(xi)(xi)(xi)(xi)板(ban)(ban)(ban)(ban)5次。操作(zuo)步驟1. 從室(shi)溫平(ping)衡20min后(hou)的(de)(de)(de)(de)鋁箔袋(dai)(dai)中(zhong)取出所(suo)需(xu)板(ban)(ban)(ban)(ban)條(tiao)，剩余板(ban)(ban)(ban)(ban)條(tiao)用(yong)(yong)(yong)(yong)自封(feng)(feng)(feng)(feng)袋(dai)(dai)密封(feng)(feng)(feng)(feng)放(fang)回(hui)4℃。2. 設置(zhi)標(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)和(he)(he)樣(yang)本(ben)(ben)孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)，標(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)各加(jia)不(bu)同濃(nong)(nong)度(du)的(de)(de)(de)(de)標(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)50μL；3. 樣(yang)本(ben)(ben)孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)先(xian)加(jia)待測樣(yang)本(ben)(ben)10μL，再加(jia)樣(yang)本(ben)(ben)稀(xi)(xi)釋(shi)(shi)液(ye)(ye)(ye)(ye)40μL；空(kong)白孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)不(bu)加(jia)。4. 除空(kong)白孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)外，標(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)和(he)(he)樣(yang)本(ben)(ben)孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)中(zhong)每孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)加(jia)入(ru)(ru)(ru)辣根過(guo)(guo)氧化(hua)(hua)物(wu)(wu)酶(mei)（HRP）標(biao)(biao)(biao)記的(de)(de)(de)(de)檢(jian)(jian)測抗(kang)體(ti)(ti)100μL，用(yong)(yong)(yong)(yong)封(feng)(feng)(feng)(feng)板(ban)(ban)(ban)(ban)膜封(feng)(feng)(feng)(feng)住反應(ying)孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)，37℃水(shui)(shui)(shui)浴(yu)鍋(guo)或恒(heng)溫箱溫育(yu)(yu)60min。5. 棄去液(ye)(ye)(ye)(ye)體(ti)(ti)，吸水(shui)(shui)(shui)紙(zhi)(zhi)上(shang)(shang)拍干，每孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)加(jia)滿洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)液(ye)(ye)(ye)(ye)，靜置(zhi)1min，甩去洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)液(ye)(ye)(ye)(ye)，吸水(shui)(shui)(shui)紙(zhi)(zhi)上(shang)(shang)拍干，如此重(zhong)復(fu)洗(xi)(xi)(xi)(xi)(xi)板(ban)(ban)(ban)(ban)5次（也可(ke)(ke)用(yong)(yong)(yong)(yong)洗(xi)(xi)(xi)(xi)(xi)板(ban)(ban)(ban)(ban)機(ji)洗(xi)(xi)(xi)(xi)(xi)板(ban)(ban)(ban)(ban)）。6. 每孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)加(jia)入(ru)(ru)(ru)底(di)物(wu)(wu)A、B各50μL，37℃避光孵育(yu)(yu)15min。7. 每孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)加(jia)入(ru)(ru)(ru)終(zhong)(zhong)止液(ye)(ye)(ye)(ye)50μL，15min內，在(zai)450nm波長(chang)處測定各孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)的(de)(de)(de)(de)OD值(zhi)。結(jie)果(guo)(guo)判斷(duan) 繪(hui)制(zhi)標(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)曲線：在(zai)Excel工作(zuo)表中(zhong)，以標(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)濃(nong)(nong)度(du)作(zuo)橫坐標(biao)(biao)(biao)，對應(ying)OD值(zhi)作(zuo)縱(zong)坐標(biao)(biao)(biao)，繪(hui)制(zhi)出標(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)線性(xing)(xing)回(hui)歸曲線，按曲線方程計算各樣(yang)本(ben)(ben)濃(nong)(nong)度(du)值(zhi)。 試(shi)(shi)(shi)劑(ji)(ji)(ji)盒(he)性(xing)(xing)能1. 準(zhun)(zhun)(zhun)(zhun)(zhun)確性(xing)(xing)：標(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)線性(xing)(xing)回(hui)歸與預期(qi)濃(nong)(nong)度(du)相關系數R值(zhi)，大(da)于(yu)等于(yu)0.9900。2. 靈(ling)敏度(du)：＊低檢(jian)(jian)測濃(nong)(nong)度(du)小于(yu)1.0 mg/mL。3. 特異性(xing)(xing)：不(bu)與其它可(ke)(ke)溶(rong)性(xing)(xing)結(jie)構(gou)類(lei)似物(wu)(wu)交叉反應(ying)。4. 重(zhong)復(fu)性(xing)(xing)：板(ban)(ban)(ban)(ban)內、板(ban)(ban)(ban)(ban)間變異系數均(jun)小于(yu)15%。5. 貯(zhu)藏：2-8℃，避光防潮保(bao)存(cun)。6. 有效(xiao)期(qi)：6個月免責聲明1. 試(shi)(shi)(shi)劑(ji)(ji)(ji)盒(he)僅供研究使用(yong)(yong)(yong)(yong)，不(bu)得用(yong)(yong)(yong)(yong)于(yu)臨床實(shi)(shi)(shi)驗(yan)(yan)或人體(ti)(ti)實(shi)(shi)(shi)驗(yan)(yan)，否則所(suo)產生的(de)(de)(de)(de)一切(qie)后(hou)果(guo)(guo)，由實(shi)(shi)(shi)驗(yan)(yan)者承擔(dan)，本(ben)(ben)公司概(gai)不(bu)負責。2. 嚴格(ge)按照(zhao)說(shuo)明書操作(zuo)，實(shi)(shi)(shi)驗(yan)(yan)者違(wei)反說(shuo)明書操作(zuo)，后(hou)果(guo)(guo)由實(shi)(shi)(shi)驗(yan)(yan)者承擔(dan)。 FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES. Human Albumin (ALB) ELISA Kit instruction Intended useThis ALB ELISA kit is intended Laboratory for Research use only and is not for use in diagnostic or therapeutic procedures.The Stop Solution changes the color from blue to yellow and the intensity of the color is measured at 450 nm using a spectrophotometer. In order to measure the concentration of ALB in the sample, this ALB ELISA Kit includes a set of calibration standards. The calibration standards are assayed at the same time as the samples and allow the operator to produce a standard curve of Optical Density versus ALB concentration. The concentration of ALB in the samples is then determined by comparing the O.D. of the samples to the standard curve.Sample collection and storagesSerum - Use a serum separator tube and allow samples to clot for 30 minutes before centrifugation for 10 minutes at approximately 3000×g. Remove serum and assay immediately or aliquot and store samples at -20℃ or -80℃.Avoid repeated freeze-thaw cyclesPlasma - Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 30 minutes at 3000×g at 2-8℃ within 30 minutes of collection. Store samples at -20℃or -80℃. Avoid repeated freeze-thaw cycles.Cell culture supernates and other biological fluids - Remove particulates by centrifugation and assay immediately or aliquot and store samples at -20℃or -80℃. Avoid repeated freeze-thaw cycles.Note: The samples shoule be centrifugated dequately and no hemolysis or granule was allowed.Materials required but not supplied1. Standard microplate reader(450nm)2. Precision pipettes and Disposable pipette tips.3. 37 ℃ incubatorPrecautions1. Do not substitute reagents from one kit to another. Standard, conjugate and microplates are matched for optimal performance. Use only the reagents supplied by manufacturer.2. Do not remove microplate from the storage bag until needed. Unused strips should be stored at 2-8°C in their pouch with the desiccant provided.3. Mix all reagents before using.Remove all kit reagents from refrigerator and allow them to reach room temperature ( 20-25°C)Materials suppliedName96 determinations48 determinationsMicroelisa stripplate12*8strips12*4stripsStandard0.3ml*6tubes0.3ml*6tubesSample Diluent6.0ml3.0mlHRP-Conjugate reagent10.0ml5.0ml20X Wash solution25ml15mlChromogen Solution A6.0ml3.0mlChromogen Solution B6.0ml3.0mlStop Solution6.0ml3.0mlClosure plate membrane22User manual11Sealed bags11Note: Standard (S0 → S5) concentration was followed by:0,6.25,12.5,25,50,100 mg/mLReagent preparation20×wash solution:Dilute with Distilled or deionized water 1:20.Assay procedure1. Prepare all reagents before starting assay procedure. It is recommended that all Standards and Samples be added in duplicate to the Microelisa Stripplate.2. Add standard: Set Standard wells, testing sample wells. Add standard 50μl to standard well.3. Add Sample: Add testing sample 10μl then add Sample Diluent 40μl to testing sample well; Blank well doesn’t add anyting.4. Add 100μl of HRP-conjugate reagent to each well, cover with an adhesive strip and incubate for 60 minutes at 37°C. 5. Aspirate each well and wash, repeating the process four times for a total of five washes. Wash by filling each well with Wash Solution (400μl) using a squirt bottle, manifold dispenser or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Solution by aspirating or decanting. Invert the plate and blot it against clean paper towels.6. Add chromogen solution A 50μl and chromogen solution B 50μl to each well. Gently mix and incubate for 15 minutes at 37°C. Protect from light.7. Add 50μl Stop Solution to each well. The color in the wells should change from blue to yellow. If the color in the wells is green or the color change does not appear uniform, gently tap the plate to ensure thorough mixing.8. Read the Optical Density (O.D.) at 450 nm using a microtiter plate reader within 15 minutes.Calculation of results1. This standard curve is used to determine the amount in an unknown sample. The standard curve is generated by plotting the average O.D. (450 nm) obtained for each of the six standard concentrations on the vertical (Y) axis versus the corresponding concentration on the horizontal (X) axis. 2. First, calculate the mean O.D. value for each standard and sample. All O.D. values, are subtracted by the mean value of the zero standard before result interpretation. Construct the standard curve using graph paper or statistical software. 3. To determine the amount in each sample, first locate the O.D. value on the Y-axis and extend a horizontal line to the standard curve. At the point of intersection, draw a vertical line to the X-axis and read the corresponding concentration. 4. Any variation in operator, pipetting and washing technique, incubation time or temperature, and kit age can cause variation in result. Each user should obtain their own standard curve.5. The sensitivity by this assay is 0.1 mg/mL6. Standard curve Storage： 2-8℃.validity： six months. FOR RESEARCH USE ONLY; NOT FOR THERAPEUTIC OR DIAGNOSTIC APPLICATIONS! PLEASE READ THROUGH ENTIRE PROCEDURE BEFORE BEGINNING!
人(ren)（Human）視(shi)神(shen)(shen)(shen)經(jing)(jing)(jing)(jing)萎(wei)縮(suo)癥(zheng)(zheng)蛋白(bai)(bai)(bai)1（OPA1） ELISA檢(jian)測(ce)(ce)(ce)(ce)(ce)(ce)試劑(ji)(ji)(ji)(ji)盒(he)(he)檢(jian)測(ce)(ce)(ce)(ce)(ce)(ce)原理試劑(ji)(ji)(ji)(ji)盒(he)(he)采用(yong)(yong)(yong)(yong)(yong)(yong)雙抗(kang)體(ti)(ti)一(yi)步夾心(xin)(xin)法酶(mei)(mei)聯免疫吸(xi)附試驗(yan)（ELISA）。往(wang)預先(xian)包被(bei)視(shi)神(shen)(shen)(shen)經(jing)(jing)(jing)(jing)萎(wei)縮(suo)癥(zheng)(zheng)蛋白(bai)(bai)(bai)1（OPA1）抗(kang)體(ti)(ti)的(de)(de)(de)包被(bei)微孔(kong)(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)(zhong)，依次加(jia)(jia)(jia)(jia)入(ru)標(biao)(biao)本(ben)、標(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)、HRP標(biao)(biao)記的(de)(de)(de)檢(jian)測(ce)(ce)(ce)(ce)(ce)(ce)抗(kang)體(ti)(ti)，經(jing)(jing)(jing)(jing)過(guo)溫育(yu)(yu)并(bing)徹底(di)洗(xi)(xi)(xi)(xi)滌(di)。用(yong)(yong)(yong)(yong)(yong)(yong)底(di)物(wu)(wu)(wu)(wu)TMB顯色，TMB在(zai)(zai)(zai)過(guo)氧(yang)化(hua)(hua)物(wu)(wu)(wu)(wu)酶(mei)(mei)的(de)(de)(de)催(cui)化(hua)(hua)下(xia)轉(zhuan)化(hua)(hua)成藍色，并(bing)在(zai)(zai)(zai)酸的(de)(de)(de)作(zuo)(zuo)用(yong)(yong)(yong)(yong)(yong)(yong)下(xia)轉(zhuan)化(hua)(hua)成＊終(zhong)(zhong)的(de)(de)(de)黃(huang)色。顏色的(de)(de)(de)深(shen)淺(qian)和(he)(he)樣(yang)(yang)品(pin)(pin)(pin)(pin)(pin)中(zhong)(zhong)(zhong)的(de)(de)(de)視(shi)神(shen)(shen)(shen)經(jing)(jing)(jing)(jing)萎(wei)縮(suo)癥(zheng)(zheng)蛋白(bai)(bai)(bai)1（OPA1）呈正(zheng)相關。用(yong)(yong)(yong)(yong)(yong)(yong)酶(mei)(mei)標(biao)(biao)儀(yi)在(zai)(zai)(zai)450nm 波長(chang)下(xia)測(ce)(ce)(ce)(ce)(ce)(ce)定(ding)吸(xi)光(guang)(guang)度(du)（OD 值(zhi)），計算(suan)(suan)樣(yang)(yang)品(pin)(pin)(pin)(pin)(pin)濃(nong)(nong)度(du)。人(ren)（Human）視(shi)神(shen)(shen)(shen)經(jing)(jing)(jing)(jing)萎(wei)縮(suo)癥(zheng)(zheng)蛋白(bai)(bai)(bai)1（OPA1） ELISA檢(jian)測(ce)(ce)(ce)(ce)(ce)(ce)試劑(ji)(ji)(ji)(ji)盒(he)(he)試劑(ji)(ji)(ji)(ji)盒(he)(he)組成名稱96孔(kong)(kong)(kong)(kong)(kong)(kong)配置48孔(kong)(kong)(kong)(kong)(kong)(kong)配置備注(zhu)(zhu)微孔(kong)(kong)(kong)(kong)(kong)(kong)酶(mei)(mei)標(biao)(biao)板(ban)12孔(kong)(kong)(kong)(kong)(kong)(kong)×8條12孔(kong)(kong)(kong)(kong)(kong)(kong)×4條無(wu)標(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)0.3mL*6管0.3mL*6管無(wu)樣(yang)(yang)本(ben)稀(xi)釋液(ye)(ye)(ye)6mL3mL無(wu)檢(jian)測(ce)(ce)(ce)(ce)(ce)(ce)抗(kang)體(ti)(ti)-HRP10mL5mL無(wu)20×洗(xi)(xi)(xi)(xi)滌(di)緩沖(chong)(chong)液(ye)(ye)(ye)25mL15mL按(an)說(shuo)明(ming)(ming)(ming)書(shu)進行稀(xi)釋底(di)物(wu)(wu)(wu)(wu)A6mL3mL無(wu)底(di)物(wu)(wu)(wu)(wu)B6mL3mL無(wu)終(zhong)(zhong)止(zhi)液(ye)(ye)(ye)6mL3mL無(wu)封(feng)(feng)(feng)板(ban)膜(mo)2張2張無(wu)說(shuo)明(ming)(ming)(ming)書(shu)1份1份無(wu)自(zi)封(feng)(feng)(feng)袋(dai)1個(ge)1個(ge)無(wu)注(zhu)(zhu)：標(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)（S0-S5）濃(nong)(nong)度(du)依次為：0、5、10、20、40、80 ng/ml樣(yang)(yang)品(pin)(pin)(pin)(pin)(pin)收集、處理及(ji)(ji)(ji)(ji)保(bao)(bao)存(cun)方法1. 血清：使(shi)用(yong)(yong)(yong)(yong)(yong)(yong)不(bu)含熱原和(he)(he)內毒素的(de)(de)(de)試管，操(cao)作(zuo)(zuo)過(guo)程(cheng)中(zhong)(zhong)(zhong)避免任(ren)何細胞刺激，收集血液(ye)(ye)(ye)后(hou)(hou)(hou)，3000轉(zhuan)離心(xin)(xin)10分(fen)(fen)(fen)鐘將(jiang)血清和(he)(he)紅細胞迅速(su)小(xiao)心(xin)(xin)地(di)(di)分(fen)(fen)(fen)離。2. 血漿：EDTA、檸檬酸鹽(yan)或(huo)肝素抗(kang)凝。3000轉(zhuan)離心(xin)(xin)30分(fen)(fen)(fen)鐘取(qu)(qu)上(shang)清。3. 細胞上(shang)清液(ye)(ye)(ye)：3000轉(zhuan)離心(xin)(xin)10分(fen)(fen)(fen)鐘去(qu)除(chu)顆(ke)粒和(he)(he)聚合物(wu)(wu)(wu)(wu)。4. 組織(zhi)勻(yun)(yun)漿：將(jiang)組織(zhi)加(jia)(jia)(jia)(jia)入(ru)適(shi)量生(sheng)(sheng)理鹽(yan)水(shui)搗碎。3000轉(zhuan)離心(xin)(xin)10分(fen)(fen)(fen)鐘取(qu)(qu)上(shang)清。5. 保(bao)(bao)存(cun)：如(ru)果(guo)樣(yang)(yang)本(ben)收集后(hou)(hou)(hou)不(bu)及(ji)(ji)(ji)(ji)時檢(jian)測(ce)(ce)(ce)(ce)(ce)(ce)，請按(an)一(yi)次用(yong)(yong)(yong)(yong)(yong)(yong)量分(fen)(fen)(fen)裝(zhuang)，凍(dong)存(cun)于(yu)-20℃，避免反(fan)復凍(dong)融，在(zai)(zai)(zai)室溫下(xia)解凍(dong)并(bing)確保(bao)(bao)樣(yang)(yang)品(pin)(pin)(pin)(pin)(pin)均(jun)勻(yun)(yun)地(di)(di)充(chong)分(fen)(fen)(fen)解凍(dong)。人(ren)（Human）視(shi)神(shen)(shen)(shen)經(jing)(jing)(jing)(jing)萎(wei)縮(suo)癥(zheng)(zheng)蛋白(bai)(bai)(bai)1（OPA1） ELISA檢(jian)測(ce)(ce)(ce)(ce)(ce)(ce)試劑(ji)(ji)(ji)(ji)盒(he)(he)自(zi)備物(wu)(wu)(wu)(wu)品(pin)(pin)(pin)(pin)(pin)1. 酶(mei)(mei)標(biao)(biao)儀(yi)（450nm）2. 高精度(du)加(jia)(jia)(jia)(jia)樣(yang)(yang)器及(ji)(ji)(ji)(ji)槍頭：0.5-10uL、2-20uL、20-200uL、200-1000uL3. 37℃恒溫箱(xiang)人(ren)（Human）視(shi)神(shen)(shen)(shen)經(jing)(jing)(jing)(jing)萎(wei)縮(suo)癥(zheng)(zheng)蛋白(bai)(bai)(bai)1（OPA1） ELISA檢(jian)測(ce)(ce)(ce)(ce)(ce)(ce)試劑(ji)(ji)(ji)(ji)盒(he)(he)操(cao)作(zuo)(zuo)注(zhu)(zhu)意事項1. 試劑(ji)(ji)(ji)(ji)盒(he)(he)保(bao)(bao)存(cun)在(zai)(zai)(zai)2-8℃，使(shi)用(yong)(yong)(yong)(yong)(yong)(yong)前室溫平衡(heng)20分(fen)(fen)(fen)鐘。從冰箱(xiang)取(qu)(qu)出(chu)(chu)的(de)(de)(de)濃(nong)(nong)縮(suo)洗(xi)(xi)(xi)(xi)滌(di)液(ye)(ye)(ye)會有結(jie)晶，這屬于(yu)正(zheng)常現(xian)象，水(shui)浴加(jia)(jia)(jia)(jia)熱使(shi)結(jie)晶完全(quan)溶(rong)解后(hou)(hou)(hou)再(zai)使(shi)用(yong)(yong)(yong)(yong)(yong)(yong)。2. 實驗(yan)中(zhong)(zhong)(zhong)不(bu)用(yong)(yong)(yong)(yong)(yong)(yong)的(de)(de)(de)板(ban)條應立即(ji)放回自(zi)封(feng)(feng)(feng)袋(dai)中(zhong)(zhong)(zhong)，密封(feng)(feng)(feng)（低溫干(gan)燥）保(bao)(bao)存(cun)。3. 濃(nong)(nong)度(du)為0的(de)(de)(de)S0號標(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)即(ji)可(ke)視(shi)為陰(yin)性(xing)(xing)對照或(huo)者(zhe)(zhe)空白(bai)(bai)(bai)；按(an)照說(shuo)明(ming)(ming)(ming)書(shu)操(cao)作(zuo)(zuo)時樣(yang)(yang)本(ben)已經(jing)(jing)(jing)(jing)稀(xi)釋5倍(bei)，＊終(zhong)(zhong)結(jie)果(guo)乘以(yi)5才是(shi)樣(yang)(yang)本(ben)實際濃(nong)(nong)度(du)。4. 嚴格(ge)(ge)按(an)照說(shuo)明(ming)(ming)(ming)書(shu)中(zhong)(zhong)(zhong)標(biao)(biao)明(ming)(ming)(ming)的(de)(de)(de)時間、加(jia)(jia)(jia)(jia)液(ye)(ye)(ye)量及(ji)(ji)(ji)(ji)順序進行溫育(yu)(yu)操(cao)作(zuo)(zuo)。5. 所有液(ye)(ye)(ye)體(ti)(ti)組分(fen)(fen)(fen)使(shi)用(yong)(yong)(yong)(yong)(yong)(yong)前充(chong)分(fen)(fen)(fen)搖勻(yun)(yun)。試劑(ji)(ji)(ji)(ji)的(de)(de)(de)準(zhun)(zhun)備 20×洗(xi)(xi)(xi)(xi)滌(di)緩沖(chong)(chong)液(ye)(ye)(ye)的(de)(de)(de)稀(xi)釋：蒸餾(liu)水(shui)按(an)1：20稀(xi)釋，即(ji)1份的(de)(de)(de)20×洗(xi)(xi)(xi)(xi)滌(di)緩沖(chong)(chong)液(ye)(ye)(ye)加(jia)(jia)(jia)(jia)19份的(de)(de)(de)蒸餾(liu)水(shui)。人(ren)（Human）視(shi)神(shen)(shen)(shen)經(jing)(jing)(jing)(jing)萎(wei)縮(suo)癥(zheng)(zheng)蛋白(bai)(bai)(bai)1（OPA1） ELISA檢(jian)測(ce)(ce)(ce)(ce)(ce)(ce)試劑(ji)(ji)(ji)(ji)盒(he)(he)洗(xi)(xi)(xi)(xi)板(ban)方法1. 手工洗(xi)(xi)(xi)(xi)板(ban)：甩(shuai)盡孔(kong)(kong)(kong)(kong)(kong)(kong)內液(ye)(ye)(ye)體(ti)(ti)，每孔(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)(jia)(jia)滿洗(xi)(xi)(xi)(xi)滌(di)液(ye)(ye)(ye)，靜(jing)置1min后(hou)(hou)(hou)甩(shuai)盡孔(kong)(kong)(kong)(kong)(kong)(kong)內液(ye)(ye)(ye)體(ti)(ti)，在(zai)(zai)(zai)吸(xi)水(shui)紙上(shang)拍(pai)(pai)干(gan)，如(ru)此洗(xi)(xi)(xi)(xi)板(ban)5次。2. 自(zi)動洗(xi)(xi)(xi)(xi)板(ban)機(ji)：每孔(kong)(kong)(kong)(kong)(kong)(kong)注(zhu)(zhu)入(ru)洗(xi)(xi)(xi)(xi)液(ye)(ye)(ye)350μL，浸泡1min，洗(xi)(xi)(xi)(xi)板(ban)5次。操(cao)作(zuo)(zuo)步驟1. 從室溫平衡(heng)20min后(hou)(hou)(hou)的(de)(de)(de)鋁(lv)箔袋(dai)中(zhong)(zhong)(zhong)取(qu)(qu)出(chu)(chu)所需(xu)板(ban)條，剩余板(ban)條用(yong)(yong)(yong)(yong)(yong)(yong)自(zi)封(feng)(feng)(feng)袋(dai)密封(feng)(feng)(feng)放回4℃。2. 設置標(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)孔(kong)(kong)(kong)(kong)(kong)(kong)和(he)(he)樣(yang)(yang)本(ben)孔(kong)(kong)(kong)(kong)(kong)(kong)，標(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)孔(kong)(kong)(kong)(kong)(kong)(kong)各(ge)(ge)加(jia)(jia)(jia)(jia)不(bu)同濃(nong)(nong)度(du)的(de)(de)(de)標(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)50μL；3. 樣(yang)(yang)本(ben)孔(kong)(kong)(kong)(kong)(kong)(kong)先(xian)加(jia)(jia)(jia)(jia)待測(ce)(ce)(ce)(ce)(ce)(ce)樣(yang)(yang)本(ben)10μL，再(zai)加(jia)(jia)(jia)(jia)樣(yang)(yang)本(ben)稀(xi)釋液(ye)(ye)(ye)40μL；空白(bai)(bai)(bai)孔(kong)(kong)(kong)(kong)(kong)(kong)不(bu)加(jia)(jia)(jia)(jia)。4. 除(chu)空白(bai)(bai)(bai)孔(kong)(kong)(kong)(kong)(kong)(kong)外，標(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)孔(kong)(kong)(kong)(kong)(kong)(kong)和(he)(he)樣(yang)(yang)本(ben)孔(kong)(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)(zhong)每孔(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)(jia)(jia)入(ru)辣根過(guo)氧(yang)化(hua)(hua)物(wu)(wu)(wu)(wu)酶(mei)(mei)（HRP）標(biao)(biao)記的(de)(de)(de)檢(jian)測(ce)(ce)(ce)(ce)(ce)(ce)抗(kang)體(ti)(ti)100μL，用(yong)(yong)(yong)(yong)(yong)(yong)封(feng)(feng)(feng)板(ban)膜(mo)封(feng)(feng)(feng)住(zhu)反(fan)應孔(kong)(kong)(kong)(kong)(kong)(kong)，37℃水(shui)浴鍋或(huo)恒溫箱(xiang)溫育(yu)(yu)60min。5. 棄去(qu)液(ye)(ye)(ye)體(ti)(ti)，吸(xi)水(shui)紙上(shang)拍(pai)(pai)干(gan)，每孔(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)(jia)(jia)滿洗(xi)(xi)(xi)(xi)滌(di)液(ye)(ye)(ye)，靜(jing)置1min，甩(shuai)去(qu)洗(xi)(xi)(xi)(xi)滌(di)液(ye)(ye)(ye)，吸(xi)水(shui)紙上(shang)拍(pai)(pai)干(gan)，如(ru)此重復洗(xi)(xi)(xi)(xi)板(ban)5次（也可(ke)用(yong)(yong)(yong)(yong)(yong)(yong)洗(xi)(xi)(xi)(xi)板(ban)機(ji)洗(xi)(xi)(xi)(xi)板(ban)）。6. 每孔(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)(jia)(jia)入(ru)底(di)物(wu)(wu)(wu)(wu)A、B各(ge)(ge)50μL，37℃避光(guang)(guang)孵(fu)育(yu)(yu)15min。7. 每孔(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)(jia)(jia)入(ru)終(zhong)(zhong)止(zhi)液(ye)(ye)(ye)50μL，15min內，在(zai)(zai)(zai)450nm波長(chang)處測(ce)(ce)(ce)(ce)(ce)(ce)定(ding)各(ge)(ge)孔(kong)(kong)(kong)(kong)(kong)(kong)的(de)(de)(de)OD值(zhi)。人(ren)（Human）視(shi)神(shen)(shen)(shen)經(jing)(jing)(jing)(jing)萎(wei)縮(suo)癥(zheng)(zheng)蛋白(bai)(bai)(bai)1（OPA1） ELISA檢(jian)測(ce)(ce)(ce)(ce)(ce)(ce)試劑(ji)(ji)(ji)(ji)盒(he)(he)結(jie)果(guo)判斷 繪制(zhi)標(biao)(biao)準(zhun)(zhun)曲線：在(zai)(zai)(zai)Excel工作(zuo)(zuo)表中(zhong)(zhong)(zhong)，以(yi)標(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)濃(nong)(nong)度(du)作(zuo)(zuo)橫坐標(biao)(biao)，對應OD值(zhi)作(zuo)(zuo)縱坐標(biao)(biao)，繪制(zhi)出(chu)(chu)標(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)線性(xing)(xing)回歸曲線，按(an)曲線方程(cheng)計算(suan)(suan)各(ge)(ge)樣(yang)(yang)本(ben)濃(nong)(nong)度(du)值(zhi)。 試劑(ji)(ji)(ji)(ji)盒(he)(he)性(xing)(xing)能(neng)1. 準(zhun)(zhun)確性(xing)(xing)：標(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)線性(xing)(xing)回歸與(yu)預期濃(nong)(nong)度(du)相關系數(shu)R值(zhi)，大于(yu)等于(yu)0.9900。2. 靈(ling)敏度(du)：＊低檢(jian)測(ce)(ce)(ce)(ce)(ce)(ce)濃(nong)(nong)度(du)小(xiao)于(yu)1.0 ng/ml。3. 特(te)異(yi)性(xing)(xing)：不(bu)與(yu)其(qi)它可(ke)溶(rong)性(xing)(xing)結(jie)構類似物(wu)(wu)(wu)(wu)交叉反(fan)應。4. 重復性(xing)(xing)：板(ban)內、板(ban)間變異(yi)系數(shu)均(jun)小(xiao)于(yu)15%。5. 貯藏：2-8℃，避光(guang)(guang)防(fang)潮保(bao)(bao)存(cun)。6. 有效期：6個(ge)月人(ren)（Human）視(shi)神(shen)(shen)(shen)經(jing)(jing)(jing)(jing)萎(wei)縮(suo)癥(zheng)(zheng)蛋白(bai)(bai)(bai)1（OPA1） ELISA檢(jian)測(ce)(ce)(ce)(ce)(ce)(ce)試劑(ji)(ji)(ji)(ji)盒(he)(he)免責(ze)聲明(ming)(ming)(ming)1. 試劑(ji)(ji)(ji)(ji)盒(he)(he)僅供研(yan)究使(shi)用(yong)(yong)(yong)(yong)(yong)(yong)，不(bu)得用(yong)(yong)(yong)(yong)(yong)(yong)于(yu)臨床實驗(yan)或(huo)人(ren)體(ti)(ti)實驗(yan)，否則(ze)所產(chan)生(sheng)(sheng)的(de)(de)(de)一(yi)切(qie)后(hou)(hou)(hou)果(guo)，由實驗(yan)者(zhe)(zhe)承(cheng)擔(dan)，本(ben)公司概不(bu)負責(ze)。2. 嚴格(ge)(ge)按(an)照說(shuo)明(ming)(ming)(ming)書(shu)操(cao)作(zuo)(zuo)，實驗(yan)者(zhe)(zhe)違反(fan)說(shuo)明(ming)(ming)(ming)書(shu)操(cao)作(zuo)(zuo)，后(hou)(hou)(hou)果(guo)由實驗(yan)者(zhe)(zhe)承(cheng)擔(dan)。
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