大(da)(da)(da)鼠白(bai)(bai)三(san)烯C4(LTC4)ELISA試(shi)(shi)劑(ji)盒(he)(he)(he)(he)大(da)(da)(da)鼠白(bai)(bai)三(san)烯C4(LTC4)ELISA試(shi)(shi)劑(ji)盒(he)(he)(he)(he)檢(jian)測(ce)(ce)(ce)原理(li)試(shi)(shi)劑(ji)盒(he)(he)(he)(he)采用(yong)(yong)(yong)雙抗(kang)(kang)(kang)體(ti)(ti)(ti)一(yi)步夾心(xin)(xin)法酶(mei)(mei)聯(lian)免(mian)疫吸(xi)(xi)附試(shi)(shi)驗(yan)（ELISA）。往預先(xian)包被(bei)活(huo)性(xing)氧簇(cu)（ROS）抗(kang)(kang)(kang)體(ti)(ti)(ti)的(de)(de)(de)(de)(de)(de)包被(bei)微(wei)(wei)孔(kong)中(zhong)，依次加(jia)(jia)(jia)(jia)入(ru)(ru)標(biao)(biao)本、標(biao)(biao)準品(pin)(pin)(pin)、HRP標(biao)(biao)記的(de)(de)(de)(de)(de)(de)檢(jian)測(ce)(ce)(ce)抗(kang)(kang)(kang)體(ti)(ti)(ti)，經過溫(wen)(wen)(wen)育(yu)并(bing)(bing)徹底(di)(di)洗(xi)(xi)滌(di)(di)。用(yong)(yong)(yong)底(di)(di)物TMB顯(xian)色(se)，TMB在(zai)(zai)(zai)(zai)過氧化(hua)物酶(mei)(mei)的(de)(de)(de)(de)(de)(de)催化(hua)下轉(zhuan)(zhuan)化(hua)成(cheng)(cheng)藍色(se)，并(bing)(bing)在(zai)(zai)(zai)(zai)酸(suan)的(de)(de)(de)(de)(de)(de)作(zuo)用(yong)(yong)(yong)下轉(zhuan)(zhuan)化(hua)成(cheng)(cheng)＊終的(de)(de)(de)(de)(de)(de)黃(huang)色(se)。顏色(se)的(de)(de)(de)(de)(de)(de)深淺和(he)(he)(he)樣(yang)(yang)品(pin)(pin)(pin)中(zhong)的(de)(de)(de)(de)(de)(de)活(huo)性(xing)氧簇(cu)（ROS）呈正相關。用(yong)(yong)(yong)酶(mei)(mei)標(biao)(biao)儀在(zai)(zai)(zai)(zai)450nm 波長(chang)下測(ce)(ce)(ce)定吸(xi)(xi)光(guang)度(du)(du)(du)(du)(du)(du)（OD 值），計算(suan)樣(yang)(yang)品(pin)(pin)(pin)活(huo)性(xing)。大(da)(da)(da)鼠白(bai)(bai)三(san)烯C4(LTC4)ELISA試(shi)(shi)劑(ji)盒(he)(he)(he)(he)樣(yang)(yang)品(pin)(pin)(pin)收(shou)集(ji)、處理(li)及保(bao)(bao)(bao)存(cun)(cun)(cun)(cun)方(fang)(fang)(fang)法1. 血(xue)清(qing)：使(shi)用(yong)(yong)(yong)不含熱原和(he)(he)(he)內毒素的(de)(de)(de)(de)(de)(de)試(shi)(shi)管(guan)，操(cao)作(zuo)過程(cheng)中(zhong)避(bi)(bi)免(mian)任何細胞刺激，收(shou)集(ji)血(xue)液(ye)(ye)后(hou)，3000轉(zhuan)(zhuan)離心(xin)(xin)10分(fen)(fen)(fen)(fen)鐘(zhong)(zhong)將血(xue)清(qing)和(he)(he)(he)紅細胞迅速(su)小心(xin)(xin)地分(fen)(fen)(fen)(fen)離。2. 血(xue)漿(jiang)(jiang)：EDTA、檸(ning)檬酸(suan)鹽(yan)或(huo)(huo)(huo)肝素抗(kang)(kang)(kang)凝。3000轉(zhuan)(zhuan)離心(xin)(xin)30分(fen)(fen)(fen)(fen)鐘(zhong)(zhong)取(qu)上清(qing)。3. 細胞上清(qing)液(ye)(ye)：3000轉(zhuan)(zhuan)離心(xin)(xin)10分(fen)(fen)(fen)(fen)鐘(zhong)(zhong)去(qu)除(chu)顆(ke)粒和(he)(he)(he)聚合物。4. 組織(zhi)勻漿(jiang)(jiang)：將組織(zhi)加(jia)(jia)(jia)(jia)入(ru)(ru)適量(liang)生理(li)鹽(yan)水搗碎。3000轉(zhuan)(zhuan)離心(xin)(xin)10分(fen)(fen)(fen)(fen)鐘(zhong)(zhong)取(qu)上清(qing)。5. 保(bao)(bao)(bao)存(cun)(cun)(cun)(cun)：如(ru)果樣(yang)(yang)本收(shou)集(ji)后(hou)不及時檢(jian)測(ce)(ce)(ce)，請按(an)一(yi)次用(yong)(yong)(yong)量(liang)分(fen)(fen)(fen)(fen)裝，凍(dong)存(cun)(cun)(cun)(cun)于(yu)-20℃，避(bi)(bi)免(mian)反復(fu)凍(dong)融(rong)，在(zai)(zai)(zai)(zai)室(shi)溫(wen)(wen)(wen)下解凍(dong)并(bing)(bing)確保(bao)(bao)(bao)樣(yang)(yang)品(pin)(pin)(pin)均勻地充分(fen)(fen)(fen)(fen)解凍(dong)。大(da)(da)(da)鼠白(bai)(bai)三(san)烯C4(LTC4)ELISA試(shi)(shi)劑(ji)盒(he)(he)(he)(he)自(zi)備(bei)物品(pin)(pin)(pin)1.酶(mei)(mei)標(biao)(biao)儀（450nm）2.高精(jing)度(du)(du)(du)(du)(du)(du)加(jia)(jia)(jia)(jia)樣(yang)(yang)器及槍(qiang)頭：0.5-10uL、2-20uL、20-200uL、200-1000uL3.37℃恒溫(wen)(wen)(wen)箱(xiang)操(cao)作(zuo)注(zhu)(zhu)意事項(xiang)1.試(shi)(shi)劑(ji)盒(he)(he)(he)(he)保(bao)(bao)(bao)存(cun)(cun)(cun)(cun)在(zai)(zai)(zai)(zai)2-8℃，使(shi)用(yong)(yong)(yong)前室(shi)溫(wen)(wen)(wen)平(ping)衡20分(fen)(fen)(fen)(fen)鐘(zhong)(zhong)。從冰箱(xiang)取(qu)出(chu)的(de)(de)(de)(de)(de)(de)濃(nong)縮洗(xi)(xi)滌(di)(di)液(ye)(ye)會有(you)結(jie)(jie)晶(jing)，這屬于(yu)正常現象，水浴(yu)加(jia)(jia)(jia)(jia)熱使(shi)結(jie)(jie)晶(jing)完全溶解后(hou)再使(shi)用(yong)(yong)(yong)。2.實(shi)驗(yan)中(zhong)不用(yong)(yong)(yong)的(de)(de)(de)(de)(de)(de)板(ban)(ban)(ban)(ban)條(tiao)(tiao)應(ying)立即放回(hui)自(zi)封(feng)(feng)(feng)袋(dai)中(zhong)，密封(feng)(feng)(feng)（低(di)(di)溫(wen)(wen)(wen)干(gan)燥）保(bao)(bao)(bao)存(cun)(cun)(cun)(cun)。3.濃(nong)度(du)(du)(du)(du)(du)(du)為(wei)0的(de)(de)(de)(de)(de)(de)S0號標(biao)(biao)準品(pin)(pin)(pin)即可視為(wei)陰性(xing)對照(zhao)或(huo)(huo)(huo)者空白(bai)(bai)；按(an)照(zhao)說(shuo)明(ming)(ming)書(shu)操(cao)作(zuo)時樣(yang)(yang)本已經稀釋(shi)5倍，＊終結(jie)(jie)果乘以5才(cai)是樣(yang)(yang)本實(shi)際濃(nong)度(du)(du)(du)(du)(du)(du)。4.嚴格(ge)按(an)照(zhao)說(shuo)明(ming)(ming)書(shu)中(zhong)標(biao)(biao)明(ming)(ming)的(de)(de)(de)(de)(de)(de)時間、加(jia)(jia)(jia)(jia)液(ye)(ye)量(liang)及順(shun)序進行(xing)溫(wen)(wen)(wen)育(yu)操(cao)作(zuo)。5.所有(you)液(ye)(ye)體(ti)(ti)(ti)組分(fen)(fen)(fen)(fen)使(shi)用(yong)(yong)(yong)前充分(fen)(fen)(fen)(fen)搖勻。試(shi)(shi)劑(ji)盒(he)(he)(he)(he)組成(cheng)(cheng)名稱(cheng)96孔(kong)配置48孔(kong)配置備(bei)注(zhu)(zhu)微(wei)(wei)孔(kong)酶(mei)(mei)標(biao)(biao)板(ban)(ban)(ban)(ban)12孔(kong)×8條(tiao)(tiao)12孔(kong)×4條(tiao)(tiao)無(wu)(wu)標(biao)(biao)準品(pin)(pin)(pin)0.3mL*6管(guan)0.3mL*6管(guan)無(wu)(wu)樣(yang)(yang)本稀釋(shi)液(ye)(ye)6mL3mL無(wu)(wu)檢(jian)測(ce)(ce)(ce)抗(kang)(kang)(kang)體(ti)(ti)(ti)-HRP10mL5mL無(wu)(wu)20×洗(xi)(xi)滌(di)(di)緩(huan)沖(chong)液(ye)(ye)25mL15mL按(an)說(shuo)明(ming)(ming)書(shu)進行(xing)稀釋(shi)底(di)(di)物A6mL3mL無(wu)(wu)底(di)(di)物B6mL3mL無(wu)(wu)終止液(ye)(ye)6mL3mL無(wu)(wu)封(feng)(feng)(feng)板(ban)(ban)(ban)(ban)膜(mo)2張2張無(wu)(wu)說(shuo)明(ming)(ming)書(shu)1份1份無(wu)(wu)自(zi)封(feng)(feng)(feng)袋(dai)1個1個無(wu)(wu)注(zhu)(zhu)：標(biao)(biao)準品(pin)(pin)(pin)（S0-S5）濃(nong)度(du)(du)(du)(du)(du)(du)依次為(wei)：0、30、60、120、240、480 U/ml試(shi)(shi)劑(ji)的(de)(de)(de)(de)(de)(de)準備(bei) 20×洗(xi)(xi)滌(di)(di)緩(huan)沖(chong)液(ye)(ye)的(de)(de)(de)(de)(de)(de)稀釋(shi)：蒸(zheng)餾水按(an)1：20稀釋(shi)，即1份的(de)(de)(de)(de)(de)(de)20×洗(xi)(xi)滌(di)(di)緩(huan)沖(chong)液(ye)(ye)加(jia)(jia)(jia)(jia)19份的(de)(de)(de)(de)(de)(de)蒸(zheng)餾水。洗(xi)(xi)板(ban)(ban)(ban)(ban)方(fang)(fang)(fang)法1.手(shou)工洗(xi)(xi)板(ban)(ban)(ban)(ban)：甩(shuai)(shuai)盡孔(kong)內液(ye)(ye)體(ti)(ti)(ti)，每(mei)孔(kong)加(jia)(jia)(jia)(jia)滿洗(xi)(xi)滌(di)(di)液(ye)(ye)，靜置1min后(hou)甩(shuai)(shuai)盡孔(kong)內液(ye)(ye)體(ti)(ti)(ti)，在(zai)(zai)(zai)(zai)吸(xi)(xi)水紙上拍(pai)干(gan)，如(ru)此洗(xi)(xi)板(ban)(ban)(ban)(ban)5次。2.自(zi)動(dong)洗(xi)(xi)板(ban)(ban)(ban)(ban)機：每(mei)孔(kong)注(zhu)(zhu)入(ru)(ru)洗(xi)(xi)液(ye)(ye)350μL，浸泡1min，洗(xi)(xi)板(ban)(ban)(ban)(ban)5次。操(cao)作(zuo)步驟1.從室(shi)溫(wen)(wen)(wen)平(ping)衡20min后(hou)的(de)(de)(de)(de)(de)(de)鋁(lv)箔(bo)袋(dai)中(zhong)取(qu)出(chu)所需(xu)板(ban)(ban)(ban)(ban)條(tiao)(tiao)，剩余板(ban)(ban)(ban)(ban)條(tiao)(tiao)用(yong)(yong)(yong)自(zi)封(feng)(feng)(feng)袋(dai)密封(feng)(feng)(feng)放回(hui)4℃。2.設置標(biao)(biao)準品(pin)(pin)(pin)孔(kong)和(he)(he)(he)樣(yang)(yang)本孔(kong)，標(biao)(biao)準品(pin)(pin)(pin)孔(kong)各(ge)(ge)加(jia)(jia)(jia)(jia)不同濃(nong)度(du)(du)(du)(du)(du)(du)的(de)(de)(de)(de)(de)(de)標(biao)(biao)準品(pin)(pin)(pin)50μL；3.樣(yang)(yang)本孔(kong)先(xian)加(jia)(jia)(jia)(jia)待(dai)測(ce)(ce)(ce)樣(yang)(yang)本10μL，再加(jia)(jia)(jia)(jia)樣(yang)(yang)本稀釋(shi)液(ye)(ye)40μL；空白(bai)(bai)孔(kong)不加(jia)(jia)(jia)(jia)。4.除(chu)空白(bai)(bai)孔(kong)外，標(biao)(biao)準品(pin)(pin)(pin)孔(kong)和(he)(he)(he)樣(yang)(yang)本孔(kong)中(zhong)每(mei)孔(kong)加(jia)(jia)(jia)(jia)入(ru)(ru)辣根過氧化(hua)物酶(mei)(mei)（HRP）標(biao)(biao)記的(de)(de)(de)(de)(de)(de)檢(jian)測(ce)(ce)(ce)抗(kang)(kang)(kang)體(ti)(ti)(ti)100μL，用(yong)(yong)(yong)封(feng)(feng)(feng)板(ban)(ban)(ban)(ban)膜(mo)封(feng)(feng)(feng)住反應(ying)孔(kong)，37℃水浴(yu)鍋(guo)或(huo)(huo)(huo)恒溫(wen)(wen)(wen)箱(xiang)溫(wen)(wen)(wen)育(yu)60min。5.棄去(qu)液(ye)(ye)體(ti)(ti)(ti)，吸(xi)(xi)水紙上拍(pai)干(gan)，每(mei)孔(kong)加(jia)(jia)(jia)(jia)滿洗(xi)(xi)滌(di)(di)液(ye)(ye)，靜置1min，甩(shuai)(shuai)去(qu)洗(xi)(xi)滌(di)(di)液(ye)(ye)，吸(xi)(xi)水紙上拍(pai)干(gan)，如(ru)此重(zhong)(zhong)復(fu)洗(xi)(xi)板(ban)(ban)(ban)(ban)5次（也可用(yong)(yong)(yong)洗(xi)(xi)板(ban)(ban)(ban)(ban)機洗(xi)(xi)板(ban)(ban)(ban)(ban)）。6.每(mei)孔(kong)加(jia)(jia)(jia)(jia)入(ru)(ru)底(di)(di)物A、B各(ge)(ge)50μL，37℃避(bi)(bi)光(guang)孵(fu)育(yu)15min。7.每(mei)孔(kong)加(jia)(jia)(jia)(jia)入(ru)(ru)終止液(ye)(ye)50μL，15min內，在(zai)(zai)(zai)(zai)450nm波長(chang)處測(ce)(ce)(ce)定各(ge)(ge)孔(kong)的(de)(de)(de)(de)(de)(de)OD值。結(jie)(jie)果判斷(duan) 繪(hui)制標(biao)(biao)準曲(qu)(qu)線：在(zai)(zai)(zai)(zai)Excel工作(zuo)表(biao)中(zhong)，以標(biao)(biao)準品(pin)(pin)(pin)濃(nong)度(du)(du)(du)(du)(du)(du)作(zuo)橫坐(zuo)標(biao)(biao)，對應(ying)OD值作(zuo)縱(zong)坐(zuo)標(biao)(biao)，繪(hui)制出(chu)標(biao)(biao)準品(pin)(pin)(pin)線性(xing)回(hui)歸曲(qu)(qu)線，按(an)曲(qu)(qu)線方(fang)(fang)(fang)程(cheng)計算(suan)各(ge)(ge)樣(yang)(yang)本濃(nong)度(du)(du)(du)(du)(du)(du)值。大(da)(da)(da)鼠白(bai)(bai)三(san)烯C4(LTC4)ELISA試(shi)(shi)劑(ji)盒(he)(he)(he)(he)試(shi)(shi)劑(ji)盒(he)(he)(he)(he)性(xing)能1.準確性(xing)：標(biao)(biao)準品(pin)(pin)(pin)線性(xing)回(hui)歸與(yu)預期(qi)濃(nong)度(du)(du)(du)(du)(du)(du)相關系數(shu)R值，大(da)(da)(da)于(yu)等于(yu)0.9900。2.靈(ling)敏度(du)(du)(du)(du)(du)(du)：＊低(di)(di)檢(jian)測(ce)(ce)(ce)濃(nong)度(du)(du)(du)(du)(du)(du)小于(yu)1.0 U/ml。3.特(te)異(yi)性(xing)：不與(yu)其它(ta)可溶性(xing)結(jie)(jie)構類似(si)物交叉反應(ying)。4.重(zhong)(zhong)復(fu)性(xing)：板(ban)(ban)(ban)(ban)內、板(ban)(ban)(ban)(ban)間變異(yi)系數(shu)均小于(yu)15%。5.貯藏：2-8℃，避(bi)(bi)光(guang)防潮保(bao)(bao)(bao)存(cun)(cun)(cun)(cun)。6.有(you)效期(qi)：6個月免(mian)責聲明(ming)(ming)1.試(shi)(shi)劑(ji)盒(he)(he)(he)(he)僅供(gong)研究使(shi)用(yong)(yong)(yong)，不得(de)用(yong)(yong)(yong)于(yu)臨床(chuang)實(shi)驗(yan)或(huo)(huo)(huo)人體(ti)(ti)(ti)實(shi)驗(yan)，否則所產生的(de)(de)(de)(de)(de)(de)一(yi)切后(hou)果，由實(shi)驗(yan)者承擔，本公司概不負責。2.嚴格(ge)按(an)照(zhao)說(shuo)明(ming)(ming)書(shu)操(cao)作(zuo)，實(shi)驗(yan)者違反說(shuo)明(ming)(ming)書(shu)操(cao)作(zuo)，后(hou)果由實(shi)驗(yan)者承擔。 大(da)(da)(da)鼠白(bai)(bai)三(san)烯C4(LTC4)ELISA試(shi)(shi)劑(ji)盒(he)(he)(he)(he)
大(da)(da)(da)鼠白(bai)血(xue)(xue)(xue)病(bing)(bing)抑(yi)(yi)制(zhi)(zhi)(zhi)因子(zi)(LIF)ELISA試(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)(he)大(da)(da)(da)鼠白(bai)血(xue)(xue)(xue)病(bing)(bing)抑(yi)(yi)制(zhi)(zhi)(zhi)因子(zi)(LIF)ELISA試(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)(he)檢測原理試(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)(he)采用(yong)(yong)(yong)(yong)雙(shuang)抗體(ti)(ti)一(yi)步夾心(xin)(xin)法(fa)酶(mei)(mei)聯免疫吸(xi)(xi)附(fu)試(shi)(shi)(shi)(shi)驗(yan)（ELISA）。往預先(xian)包被(bei)活性氧(yang)(yang)簇（ROS）抗體(ti)(ti)的(de)(de)(de)(de)(de)(de)包被(bei)微(wei)孔(kong)(kong)中(zhong)(zhong)(zhong)(zhong)(zhong)，依(yi)次加(jia)入(ru)標本(ben)(ben)、標準品(pin)(pin)(pin)(pin)(pin)(pin)、HRP標記(ji)的(de)(de)(de)(de)(de)(de)檢測抗體(ti)(ti)，經過(guo)(guo)溫(wen)(wen)(wen)(wen)育(yu)(yu)并(bing)徹底(di)洗(xi)(xi)(xi)(xi)滌(di)(di)(di)。用(yong)(yong)(yong)(yong)底(di)物(wu)(wu)(wu)TMB顯(xian)色，TMB在(zai)(zai)過(guo)(guo)氧(yang)(yang)化(hua)(hua)物(wu)(wu)(wu)酶(mei)(mei)的(de)(de)(de)(de)(de)(de)催化(hua)(hua)下(xia)轉(zhuan)(zhuan)化(hua)(hua)成(cheng)藍色，并(bing)在(zai)(zai)酸(suan)的(de)(de)(de)(de)(de)(de)作(zuo)(zuo)(zuo)用(yong)(yong)(yong)(yong)下(xia)轉(zhuan)(zhuan)化(hua)(hua)成(cheng)＊終(zhong)的(de)(de)(de)(de)(de)(de)黃色。顏色的(de)(de)(de)(de)(de)(de)深(shen)淺和(he)(he)樣(yang)(yang)(yang)品(pin)(pin)(pin)(pin)(pin)(pin)中(zhong)(zhong)(zhong)(zhong)(zhong)的(de)(de)(de)(de)(de)(de)活性氧(yang)(yang)簇（ROS）呈正相(xiang)關(guan)(guan)。用(yong)(yong)(yong)(yong)酶(mei)(mei)標儀(yi)在(zai)(zai)450nm 波長下(xia)測定吸(xi)(xi)光度(du)(du)(du)（OD 值(zhi)），計算樣(yang)(yang)(yang)品(pin)(pin)(pin)(pin)(pin)(pin)活性。大(da)(da)(da)鼠白(bai)血(xue)(xue)(xue)病(bing)(bing)抑(yi)(yi)制(zhi)(zhi)(zhi)因子(zi)(LIF)ELISA試(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)(he)樣(yang)(yang)(yang)品(pin)(pin)(pin)(pin)(pin)(pin)收(shou)集、處理及(ji)(ji)保(bao)存方法(fa)1. 血(xue)(xue)(xue)清：使(shi)(shi)用(yong)(yong)(yong)(yong)不(bu)含熱原和(he)(he)內毒素的(de)(de)(de)(de)(de)(de)試(shi)(shi)(shi)(shi)管(guan)，操(cao)(cao)作(zuo)(zuo)(zuo)過(guo)(guo)程中(zhong)(zhong)(zhong)(zhong)(zhong)避免任何(he)細胞(bao)刺激，收(shou)集血(xue)(xue)(xue)液(ye)(ye)(ye)(ye)后(hou)(hou)(hou)，3000轉(zhuan)(zhuan)離(li)(li)心(xin)(xin)10分(fen)(fen)(fen)鐘(zhong)(zhong)將(jiang)血(xue)(xue)(xue)清和(he)(he)紅(hong)細胞(bao)迅速小心(xin)(xin)地分(fen)(fen)(fen)離(li)(li)。2. 血(xue)(xue)(xue)漿(jiang)(jiang)：EDTA、檸檬酸(suan)鹽或(huo)肝素抗凝。3000轉(zhuan)(zhuan)離(li)(li)心(xin)(xin)30分(fen)(fen)(fen)鐘(zhong)(zhong)取上(shang)清。3. 細胞(bao)上(shang)清液(ye)(ye)(ye)(ye)：3000轉(zhuan)(zhuan)離(li)(li)心(xin)(xin)10分(fen)(fen)(fen)鐘(zhong)(zhong)去除顆粒和(he)(he)聚合物(wu)(wu)(wu)。4. 組織(zhi)勻漿(jiang)(jiang)：將(jiang)組織(zhi)加(jia)入(ru)適量(liang)生(sheng)理鹽水(shui)(shui)搗碎。3000轉(zhuan)(zhuan)離(li)(li)心(xin)(xin)10分(fen)(fen)(fen)鐘(zhong)(zhong)取上(shang)清。5. 保(bao)存：如(ru)果(guo)樣(yang)(yang)(yang)本(ben)(ben)收(shou)集后(hou)(hou)(hou)不(bu)及(ji)(ji)時(shi)檢測，請按(an)一(yi)次用(yong)(yong)(yong)(yong)量(liang)分(fen)(fen)(fen)裝，凍存于-20℃，避免反復(fu)凍融，在(zai)(zai)室溫(wen)(wen)(wen)(wen)下(xia)解凍并(bing)確(que)保(bao)樣(yang)(yang)(yang)品(pin)(pin)(pin)(pin)(pin)(pin)均勻地充分(fen)(fen)(fen)解凍。大(da)(da)(da)鼠白(bai)血(xue)(xue)(xue)病(bing)(bing)抑(yi)(yi)制(zhi)(zhi)(zhi)因子(zi)(LIF)ELISA試(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)(he)自(zi)備物(wu)(wu)(wu)品(pin)(pin)(pin)(pin)(pin)(pin)1.酶(mei)(mei)標儀(yi)（450nm）2.高精度(du)(du)(du)加(jia)樣(yang)(yang)(yang)器及(ji)(ji)槍頭(tou)：0.5-10uL、2-20uL、20-200uL、200-1000uL3.37℃恒溫(wen)(wen)(wen)(wen)箱操(cao)(cao)作(zuo)(zuo)(zuo)注(zhu)意事項1.試(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)(he)保(bao)存在(zai)(zai)2-8℃，使(shi)(shi)用(yong)(yong)(yong)(yong)前室溫(wen)(wen)(wen)(wen)平衡20分(fen)(fen)(fen)鐘(zhong)(zhong)。從(cong)冰(bing)箱取出(chu)的(de)(de)(de)(de)(de)(de)濃(nong)縮洗(xi)(xi)(xi)(xi)滌(di)(di)(di)液(ye)(ye)(ye)(ye)會有結(jie)(jie)晶，這屬于正常(chang)現象，水(shui)(shui)浴加(jia)熱使(shi)(shi)結(jie)(jie)晶完全溶(rong)解后(hou)(hou)(hou)再使(shi)(shi)用(yong)(yong)(yong)(yong)。2.實(shi)驗(yan)中(zhong)(zhong)(zhong)(zhong)(zhong)不(bu)用(yong)(yong)(yong)(yong)的(de)(de)(de)(de)(de)(de)板(ban)(ban)條(tiao)應(ying)(ying)立即放(fang)回(hui)自(zi)封(feng)(feng)袋中(zhong)(zhong)(zhong)(zhong)(zhong)，密封(feng)(feng)（低溫(wen)(wen)(wen)(wen)干燥）保(bao)存。3.濃(nong)度(du)(du)(du)為0的(de)(de)(de)(de)(de)(de)S0號標準品(pin)(pin)(pin)(pin)(pin)(pin)即可(ke)視為陰性對照或(huo)者空(kong)白(bai)；按(an)照說(shuo)(shuo)明(ming)(ming)書(shu)(shu)(shu)操(cao)(cao)作(zuo)(zuo)(zuo)時(shi)樣(yang)(yang)(yang)本(ben)(ben)已經稀(xi)(xi)釋(shi)(shi)5倍，＊終(zhong)結(jie)(jie)果(guo)乘以(yi)5才是(shi)樣(yang)(yang)(yang)本(ben)(ben)實(shi)際濃(nong)度(du)(du)(du)。4.嚴格按(an)照說(shuo)(shuo)明(ming)(ming)書(shu)(shu)(shu)中(zhong)(zhong)(zhong)(zhong)(zhong)標明(ming)(ming)的(de)(de)(de)(de)(de)(de)時(shi)間、加(jia)液(ye)(ye)(ye)(ye)量(liang)及(ji)(ji)順序進(jin)行(xing)溫(wen)(wen)(wen)(wen)育(yu)(yu)操(cao)(cao)作(zuo)(zuo)(zuo)。5.所(suo)有液(ye)(ye)(ye)(ye)體(ti)(ti)組分(fen)(fen)(fen)使(shi)(shi)用(yong)(yong)(yong)(yong)前充分(fen)(fen)(fen)搖勻。試(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)(he)組成(cheng)名(ming)稱96孔(kong)(kong)配(pei)置(zhi)(zhi)48孔(kong)(kong)配(pei)置(zhi)(zhi)備注(zhu)微(wei)孔(kong)(kong)酶(mei)(mei)標板(ban)(ban)12孔(kong)(kong)×8條(tiao)12孔(kong)(kong)×4條(tiao)無(wu)標準品(pin)(pin)(pin)(pin)(pin)(pin)0.3mL*6管(guan)0.3mL*6管(guan)無(wu)樣(yang)(yang)(yang)本(ben)(ben)稀(xi)(xi)釋(shi)(shi)液(ye)(ye)(ye)(ye)6mL3mL無(wu)檢測抗體(ti)(ti)-HRP10mL5mL無(wu)20×洗(xi)(xi)(xi)(xi)滌(di)(di)(di)緩(huan)沖液(ye)(ye)(ye)(ye)25mL15mL按(an)說(shuo)(shuo)明(ming)(ming)書(shu)(shu)(shu)進(jin)行(xing)稀(xi)(xi)釋(shi)(shi)底(di)物(wu)(wu)(wu)A6mL3mL無(wu)底(di)物(wu)(wu)(wu)B6mL3mL無(wu)終(zhong)止(zhi)液(ye)(ye)(ye)(ye)6mL3mL無(wu)封(feng)(feng)板(ban)(ban)膜(mo)(mo)2張2張無(wu)說(shuo)(shuo)明(ming)(ming)書(shu)(shu)(shu)1份(fen)1份(fen)無(wu)自(zi)封(feng)(feng)袋1個1個無(wu)注(zhu)：標準品(pin)(pin)(pin)(pin)(pin)(pin)（S0-S5）濃(nong)度(du)(du)(du)依(yi)次為：0、30、60、120、240、480 U/ml試(shi)(shi)(shi)(shi)劑(ji)(ji)的(de)(de)(de)(de)(de)(de)準備 20×洗(xi)(xi)(xi)(xi)滌(di)(di)(di)緩(huan)沖液(ye)(ye)(ye)(ye)的(de)(de)(de)(de)(de)(de)稀(xi)(xi)釋(shi)(shi)：蒸餾(liu)水(shui)(shui)按(an)1：20稀(xi)(xi)釋(shi)(shi)，即1份(fen)的(de)(de)(de)(de)(de)(de)20×洗(xi)(xi)(xi)(xi)滌(di)(di)(di)緩(huan)沖液(ye)(ye)(ye)(ye)加(jia)19份(fen)的(de)(de)(de)(de)(de)(de)蒸餾(liu)水(shui)(shui)。洗(xi)(xi)(xi)(xi)板(ban)(ban)方法(fa)1.手工洗(xi)(xi)(xi)(xi)板(ban)(ban)：甩盡孔(kong)(kong)內液(ye)(ye)(ye)(ye)體(ti)(ti)，每孔(kong)(kong)加(jia)滿洗(xi)(xi)(xi)(xi)滌(di)(di)(di)液(ye)(ye)(ye)(ye)，靜置(zhi)(zhi)1min后(hou)(hou)(hou)甩盡孔(kong)(kong)內液(ye)(ye)(ye)(ye)體(ti)(ti)，在(zai)(zai)吸(xi)(xi)水(shui)(shui)紙(zhi)上(shang)拍(pai)(pai)干，如(ru)此(ci)洗(xi)(xi)(xi)(xi)板(ban)(ban)5次。2.自(zi)動洗(xi)(xi)(xi)(xi)板(ban)(ban)機(ji)：每孔(kong)(kong)注(zhu)入(ru)洗(xi)(xi)(xi)(xi)液(ye)(ye)(ye)(ye)350μL，浸泡1min，洗(xi)(xi)(xi)(xi)板(ban)(ban)5次。操(cao)(cao)作(zuo)(zuo)(zuo)步驟(zou)1.從(cong)室溫(wen)(wen)(wen)(wen)平衡20min后(hou)(hou)(hou)的(de)(de)(de)(de)(de)(de)鋁箔袋中(zhong)(zhong)(zhong)(zhong)(zhong)取出(chu)所(suo)需(xu)板(ban)(ban)條(tiao)，剩余(yu)板(ban)(ban)條(tiao)用(yong)(yong)(yong)(yong)自(zi)封(feng)(feng)袋密封(feng)(feng)放(fang)回(hui)4℃。2.設置(zhi)(zhi)標準品(pin)(pin)(pin)(pin)(pin)(pin)孔(kong)(kong)和(he)(he)樣(yang)(yang)(yang)本(ben)(ben)孔(kong)(kong)，標準品(pin)(pin)(pin)(pin)(pin)(pin)孔(kong)(kong)各加(jia)不(bu)同(tong)濃(nong)度(du)(du)(du)的(de)(de)(de)(de)(de)(de)標準品(pin)(pin)(pin)(pin)(pin)(pin)50μL；3.樣(yang)(yang)(yang)本(ben)(ben)孔(kong)(kong)先(xian)加(jia)待測樣(yang)(yang)(yang)本(ben)(ben)10μL，再加(jia)樣(yang)(yang)(yang)本(ben)(ben)稀(xi)(xi)釋(shi)(shi)液(ye)(ye)(ye)(ye)40μL；空(kong)白(bai)孔(kong)(kong)不(bu)加(jia)。4.除空(kong)白(bai)孔(kong)(kong)外，標準品(pin)(pin)(pin)(pin)(pin)(pin)孔(kong)(kong)和(he)(he)樣(yang)(yang)(yang)本(ben)(ben)孔(kong)(kong)中(zhong)(zhong)(zhong)(zhong)(zhong)每孔(kong)(kong)加(jia)入(ru)辣根(gen)過(guo)(guo)氧(yang)(yang)化(hua)(hua)物(wu)(wu)(wu)酶(mei)(mei)（HRP）標記(ji)的(de)(de)(de)(de)(de)(de)檢測抗體(ti)(ti)100μL，用(yong)(yong)(yong)(yong)封(feng)(feng)板(ban)(ban)膜(mo)(mo)封(feng)(feng)住反應(ying)(ying)孔(kong)(kong)，37℃水(shui)(shui)浴鍋(guo)或(huo)恒溫(wen)(wen)(wen)(wen)箱溫(wen)(wen)(wen)(wen)育(yu)(yu)60min。5.棄去液(ye)(ye)(ye)(ye)體(ti)(ti)，吸(xi)(xi)水(shui)(shui)紙(zhi)上(shang)拍(pai)(pai)干，每孔(kong)(kong)加(jia)滿洗(xi)(xi)(xi)(xi)滌(di)(di)(di)液(ye)(ye)(ye)(ye)，靜置(zhi)(zhi)1min，甩去洗(xi)(xi)(xi)(xi)滌(di)(di)(di)液(ye)(ye)(ye)(ye)，吸(xi)(xi)水(shui)(shui)紙(zhi)上(shang)拍(pai)(pai)干，如(ru)此(ci)重(zhong)復(fu)洗(xi)(xi)(xi)(xi)板(ban)(ban)5次（也可(ke)用(yong)(yong)(yong)(yong)洗(xi)(xi)(xi)(xi)板(ban)(ban)機(ji)洗(xi)(xi)(xi)(xi)板(ban)(ban)）。6.每孔(kong)(kong)加(jia)入(ru)底(di)物(wu)(wu)(wu)A、B各50μL，37℃避光孵育(yu)(yu)15min。7.每孔(kong)(kong)加(jia)入(ru)終(zhong)止(zhi)液(ye)(ye)(ye)(ye)50μL，15min內，在(zai)(zai)450nm波長處測定各孔(kong)(kong)的(de)(de)(de)(de)(de)(de)OD值(zhi)。結(jie)(jie)果(guo)判(pan)斷 繪制(zhi)(zhi)(zhi)標準曲線(xian)：在(zai)(zai)Excel工作(zuo)(zuo)(zuo)表(biao)中(zhong)(zhong)(zhong)(zhong)(zhong)，以(yi)標準品(pin)(pin)(pin)(pin)(pin)(pin)濃(nong)度(du)(du)(du)作(zuo)(zuo)(zuo)橫坐標，對應(ying)(ying)OD值(zhi)作(zuo)(zuo)(zuo)縱坐標，繪制(zhi)(zhi)(zhi)出(chu)標準品(pin)(pin)(pin)(pin)(pin)(pin)線(xian)性回(hui)歸曲線(xian)，按(an)曲線(xian)方程計算各樣(yang)(yang)(yang)本(ben)(ben)濃(nong)度(du)(du)(du)值(zhi)。大(da)(da)(da)鼠白(bai)血(xue)(xue)(xue)病(bing)(bing)抑(yi)(yi)制(zhi)(zhi)(zhi)因子(zi)(LIF)ELISA試(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)(he)試(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)(he)性能1.準確(que)性：標準品(pin)(pin)(pin)(pin)(pin)(pin)線(xian)性回(hui)歸與(yu)預期濃(nong)度(du)(du)(du)相(xiang)關(guan)(guan)系(xi)數R值(zhi)，大(da)(da)(da)于等于0.9900。2.靈敏度(du)(du)(du)：＊低檢測濃(nong)度(du)(du)(du)小于1.0 U/ml。3.特(te)異性：不(bu)與(yu)其它(ta)可(ke)溶(rong)性結(jie)(jie)構(gou)類似物(wu)(wu)(wu)交(jiao)叉反應(ying)(ying)。4.重(zhong)復(fu)性：板(ban)(ban)內、板(ban)(ban)間變異系(xi)數均小于15%。5.貯藏：2-8℃，避光防(fang)潮保(bao)存。6.有效期：6個月免責聲明(ming)(ming)1.試(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)(he)僅供研究使(shi)(shi)用(yong)(yong)(yong)(yong)，不(bu)得用(yong)(yong)(yong)(yong)于臨床實(shi)驗(yan)或(huo)人體(ti)(ti)實(shi)驗(yan)，否(fou)則所(suo)產生(sheng)的(de)(de)(de)(de)(de)(de)一(yi)切后(hou)(hou)(hou)果(guo)，由實(shi)驗(yan)者承(cheng)擔，本(ben)(ben)公司概(gai)不(bu)負責。2.嚴格按(an)照說(shuo)(shuo)明(ming)(ming)書(shu)(shu)(shu)操(cao)(cao)作(zuo)(zuo)(zuo)，實(shi)驗(yan)者違反說(shuo)(shuo)明(ming)(ming)書(shu)(shu)(shu)操(cao)(cao)作(zuo)(zuo)(zuo)，后(hou)(hou)(hou)果(guo)由實(shi)驗(yan)者承(cheng)擔。 大(da)(da)(da)鼠白(bai)血(xue)(xue)(xue)病(bing)(bing)抑(yi)(yi)制(zhi)(zhi)(zhi)因子(zi)(LIF)ELISA試(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)(he)
大(da)鼠白(bai)(bai)血(xue)(xue)(xue)病(bing)(bing)抑制因(yin)(yin)子(zi)(zi)受體(ti)(ti)(ti)(ti)(ti)(ti)(ti)(LIFR)ELISA試(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)大(da)鼠白(bai)(bai)血(xue)(xue)(xue)病(bing)(bing)抑制因(yin)(yin)子(zi)(zi)受體(ti)(ti)(ti)(ti)(ti)(ti)(ti)(LIFR)ELISA試(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)檢(jian)測(ce)原理試(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)采用(yong)(yong)(yong)雙抗(kang)體(ti)(ti)(ti)(ti)(ti)(ti)(ti)一步夾心法(fa)酶聯免(mian)(mian)疫吸(xi)附試(shi)(shi)(shi)(shi)驗(yan)（ELISA）。往(wang)預先包被活(huo)(huo)性氧(yang)簇（ROS）抗(kang)體(ti)(ti)(ti)(ti)(ti)(ti)(ti)的(de)包被微孔(kong)(kong)(kong)(kong)中(zhong)，依(yi)次(ci)加(jia)(jia)(jia)入(ru)標(biao)(biao)(biao)(biao)本、標(biao)(biao)(biao)(biao)準(zhun)品(pin)(pin)(pin)、HRP標(biao)(biao)(biao)(biao)記的(de)檢(jian)測(ce)抗(kang)體(ti)(ti)(ti)(ti)(ti)(ti)(ti)，經過(guo)溫育(yu)(yu)并(bing)徹(che)底洗(xi)(xi)(xi)滌(di)。用(yong)(yong)(yong)底物(wu)TMB顯色(se)，TMB在(zai)過(guo)氧(yang)化(hua)(hua)物(wu)酶的(de)催化(hua)(hua)下(xia)轉化(hua)(hua)成(cheng)藍色(se)，并(bing)在(zai)酸(suan)的(de)作(zuo)(zuo)用(yong)(yong)(yong)下(xia)轉化(hua)(hua)成(cheng)＊終(zhong)的(de)黃色(se)。顏色(se)的(de)深(shen)淺和(he)樣品(pin)(pin)(pin)中(zhong)的(de)活(huo)(huo)性氧(yang)簇（ROS）呈正(zheng)(zheng)相關。用(yong)(yong)(yong)酶標(biao)(biao)(biao)(biao)儀在(zai)450nm 波長(chang)下(xia)測(ce)定(ding)吸(xi)光度(du)(du)（OD 值(zhi)），計(ji)(ji)算(suan)樣品(pin)(pin)(pin)活(huo)(huo)性。大(da)鼠白(bai)(bai)血(xue)(xue)(xue)病(bing)(bing)抑制因(yin)(yin)子(zi)(zi)受體(ti)(ti)(ti)(ti)(ti)(ti)(ti)(LIFR)ELISA試(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)樣品(pin)(pin)(pin)收(shou)集(ji)、處(chu)理及(ji)保(bao)(bao)存(cun)方(fang)法(fa)1. 血(xue)(xue)(xue)清：使用(yong)(yong)(yong)不(bu)(bu)(bu)含熱(re)原和(he)內(nei)毒素的(de)試(shi)(shi)(shi)(shi)管，操(cao)(cao)(cao)作(zuo)(zuo)過(guo)程中(zhong)避(bi)免(mian)(mian)任(ren)何(he)細胞(bao)刺激，收(shou)集(ji)血(xue)(xue)(xue)液(ye)(ye)(ye)(ye)(ye)(ye)后(hou)，3000轉離(li)(li)心10分(fen)鐘(zhong)將(jiang)血(xue)(xue)(xue)清和(he)紅細胞(bao)迅速小心地分(fen)離(li)(li)。2. 血(xue)(xue)(xue)漿：EDTA、檸檬(meng)酸(suan)鹽或(huo)肝素抗(kang)凝(ning)。3000轉離(li)(li)心30分(fen)鐘(zhong)取上(shang)(shang)清。3. 細胞(bao)上(shang)(shang)清液(ye)(ye)(ye)(ye)(ye)(ye)：3000轉離(li)(li)心10分(fen)鐘(zhong)去(qu)除顆粒和(he)聚合物(wu)。4. 組(zu)織勻漿：將(jiang)組(zu)織加(jia)(jia)(jia)入(ru)適(shi)量(liang)生(sheng)理鹽水搗碎。3000轉離(li)(li)心10分(fen)鐘(zhong)取上(shang)(shang)清。5. 保(bao)(bao)存(cun)：如果樣本收(shou)集(ji)后(hou)不(bu)(bu)(bu)及(ji)時(shi)檢(jian)測(ce)，請按(an)(an)一次(ci)用(yong)(yong)(yong)量(liang)分(fen)裝，凍存(cun)于(yu)-20℃，避(bi)免(mian)(mian)反(fan)(fan)復(fu)凍融，在(zai)室溫下(xia)解(jie)凍并(bing)確(que)(que)保(bao)(bao)樣品(pin)(pin)(pin)均勻地充分(fen)解(jie)凍。大(da)鼠白(bai)(bai)血(xue)(xue)(xue)病(bing)(bing)抑制因(yin)(yin)子(zi)(zi)受體(ti)(ti)(ti)(ti)(ti)(ti)(ti)(LIFR)ELISA試(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)自(zi)備物(wu)品(pin)(pin)(pin)1.酶標(biao)(biao)(biao)(biao)儀（450nm）2.高精度(du)(du)加(jia)(jia)(jia)樣器及(ji)槍頭：0.5-10uL、2-20uL、20-200uL、200-1000uL3.37℃恒(heng)溫箱(xiang)操(cao)(cao)(cao)作(zuo)(zuo)注(zhu)意事項(xiang)1.試(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)保(bao)(bao)存(cun)在(zai)2-8℃，使用(yong)(yong)(yong)前室溫平衡20分(fen)鐘(zhong)。從冰箱(xiang)取出(chu)的(de)濃(nong)(nong)縮洗(xi)(xi)(xi)滌(di)液(ye)(ye)(ye)(ye)(ye)(ye)會有(you)結晶，這屬(shu)于(yu)正(zheng)(zheng)常現象，水浴加(jia)(jia)(jia)熱(re)使結晶完全溶解(jie)后(hou)再(zai)使用(yong)(yong)(yong)。2.實(shi)(shi)驗(yan)中(zhong)不(bu)(bu)(bu)用(yong)(yong)(yong)的(de)板(ban)(ban)(ban)條應立即放(fang)(fang)回(hui)(hui)自(zi)封(feng)袋中(zhong)，密封(feng)（低溫干(gan)燥(zao)）保(bao)(bao)存(cun)。3.濃(nong)(nong)度(du)(du)為(wei)0的(de)S0號(hao)標(biao)(biao)(biao)(biao)準(zhun)品(pin)(pin)(pin)即可視為(wei)陰性對照或(huo)者(zhe)空白(bai)(bai)；按(an)(an)照說(shuo)(shuo)明(ming)書(shu)(shu)操(cao)(cao)(cao)作(zuo)(zuo)時(shi)樣本已(yi)經稀(xi)釋5倍(bei)，＊終(zhong)結果乘以(yi)5才是樣本實(shi)(shi)際濃(nong)(nong)度(du)(du)。4.嚴格(ge)按(an)(an)照說(shuo)(shuo)明(ming)書(shu)(shu)中(zhong)標(biao)(biao)(biao)(biao)明(ming)的(de)時(shi)間、加(jia)(jia)(jia)液(ye)(ye)(ye)(ye)(ye)(ye)量(liang)及(ji)順序進行溫育(yu)(yu)操(cao)(cao)(cao)作(zuo)(zuo)。5.所(suo)有(you)液(ye)(ye)(ye)(ye)(ye)(ye)體(ti)(ti)(ti)(ti)(ti)(ti)(ti)組(zu)分(fen)使用(yong)(yong)(yong)前充分(fen)搖勻。試(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)組(zu)成(cheng)名稱96孔(kong)(kong)(kong)(kong)配置48孔(kong)(kong)(kong)(kong)配置備注(zhu)微孔(kong)(kong)(kong)(kong)酶標(biao)(biao)(biao)(biao)板(ban)(ban)(ban)12孔(kong)(kong)(kong)(kong)×8條12孔(kong)(kong)(kong)(kong)×4條無(wu)標(biao)(biao)(biao)(biao)準(zhun)品(pin)(pin)(pin)0.3mL*6管0.3mL*6管無(wu)樣本稀(xi)釋液(ye)(ye)(ye)(ye)(ye)(ye)6mL3mL無(wu)檢(jian)測(ce)抗(kang)體(ti)(ti)(ti)(ti)(ti)(ti)(ti)-HRP10mL5mL無(wu)20×洗(xi)(xi)(xi)滌(di)緩沖(chong)液(ye)(ye)(ye)(ye)(ye)(ye)25mL15mL按(an)(an)說(shuo)(shuo)明(ming)書(shu)(shu)進行稀(xi)釋底物(wu)A6mL3mL無(wu)底物(wu)B6mL3mL無(wu)終(zhong)止液(ye)(ye)(ye)(ye)(ye)(ye)6mL3mL無(wu)封(feng)板(ban)(ban)(ban)膜(mo)2張2張無(wu)說(shuo)(shuo)明(ming)書(shu)(shu)1份1份無(wu)自(zi)封(feng)袋1個1個無(wu)注(zhu)：標(biao)(biao)(biao)(biao)準(zhun)品(pin)(pin)(pin)（S0-S5）濃(nong)(nong)度(du)(du)依(yi)次(ci)為(wei)：0、30、60、120、240、480 U/ml試(shi)(shi)(shi)(shi)劑(ji)(ji)的(de)準(zhun)備 20×洗(xi)(xi)(xi)滌(di)緩沖(chong)液(ye)(ye)(ye)(ye)(ye)(ye)的(de)稀(xi)釋：蒸(zheng)餾水按(an)(an)1：20稀(xi)釋，即1份的(de)20×洗(xi)(xi)(xi)滌(di)緩沖(chong)液(ye)(ye)(ye)(ye)(ye)(ye)加(jia)(jia)(jia)19份的(de)蒸(zheng)餾水。洗(xi)(xi)(xi)板(ban)(ban)(ban)方(fang)法(fa)1.手工洗(xi)(xi)(xi)板(ban)(ban)(ban)：甩(shuai)盡孔(kong)(kong)(kong)(kong)內(nei)液(ye)(ye)(ye)(ye)(ye)(ye)體(ti)(ti)(ti)(ti)(ti)(ti)(ti)，每孔(kong)(kong)(kong)(kong)加(jia)(jia)(jia)滿洗(xi)(xi)(xi)滌(di)液(ye)(ye)(ye)(ye)(ye)(ye)，靜置1min后(hou)甩(shuai)盡孔(kong)(kong)(kong)(kong)內(nei)液(ye)(ye)(ye)(ye)(ye)(ye)體(ti)(ti)(ti)(ti)(ti)(ti)(ti)，在(zai)吸(xi)水紙(zhi)上(shang)(shang)拍(pai)干(gan)，如此洗(xi)(xi)(xi)板(ban)(ban)(ban)5次(ci)。2.自(zi)動洗(xi)(xi)(xi)板(ban)(ban)(ban)機：每孔(kong)(kong)(kong)(kong)注(zhu)入(ru)洗(xi)(xi)(xi)液(ye)(ye)(ye)(ye)(ye)(ye)350μL，浸泡1min，洗(xi)(xi)(xi)板(ban)(ban)(ban)5次(ci)。操(cao)(cao)(cao)作(zuo)(zuo)步驟1.從室溫平衡20min后(hou)的(de)鋁箔(bo)袋中(zhong)取出(chu)所(suo)需板(ban)(ban)(ban)條，剩余(yu)板(ban)(ban)(ban)條用(yong)(yong)(yong)自(zi)封(feng)袋密封(feng)放(fang)(fang)回(hui)(hui)4℃。2.設置標(biao)(biao)(biao)(biao)準(zhun)品(pin)(pin)(pin)孔(kong)(kong)(kong)(kong)和(he)樣本孔(kong)(kong)(kong)(kong)，標(biao)(biao)(biao)(biao)準(zhun)品(pin)(pin)(pin)孔(kong)(kong)(kong)(kong)各加(jia)(jia)(jia)不(bu)(bu)(bu)同濃(nong)(nong)度(du)(du)的(de)標(biao)(biao)(biao)(biao)準(zhun)品(pin)(pin)(pin)50μL；3.樣本孔(kong)(kong)(kong)(kong)先加(jia)(jia)(jia)待測(ce)樣本10μL，再(zai)加(jia)(jia)(jia)樣本稀(xi)釋液(ye)(ye)(ye)(ye)(ye)(ye)40μL；空白(bai)(bai)孔(kong)(kong)(kong)(kong)不(bu)(bu)(bu)加(jia)(jia)(jia)。4.除空白(bai)(bai)孔(kong)(kong)(kong)(kong)外，標(biao)(biao)(biao)(biao)準(zhun)品(pin)(pin)(pin)孔(kong)(kong)(kong)(kong)和(he)樣本孔(kong)(kong)(kong)(kong)中(zhong)每孔(kong)(kong)(kong)(kong)加(jia)(jia)(jia)入(ru)辣根過(guo)氧(yang)化(hua)(hua)物(wu)酶（HRP）標(biao)(biao)(biao)(biao)記的(de)檢(jian)測(ce)抗(kang)體(ti)(ti)(ti)(ti)(ti)(ti)(ti)100μL，用(yong)(yong)(yong)封(feng)板(ban)(ban)(ban)膜(mo)封(feng)住(zhu)反(fan)(fan)應孔(kong)(kong)(kong)(kong)，37℃水浴鍋或(huo)恒(heng)溫箱(xiang)溫育(yu)(yu)60min。5.棄去(qu)液(ye)(ye)(ye)(ye)(ye)(ye)體(ti)(ti)(ti)(ti)(ti)(ti)(ti)，吸(xi)水紙(zhi)上(shang)(shang)拍(pai)干(gan)，每孔(kong)(kong)(kong)(kong)加(jia)(jia)(jia)滿洗(xi)(xi)(xi)滌(di)液(ye)(ye)(ye)(ye)(ye)(ye)，靜置1min，甩(shuai)去(qu)洗(xi)(xi)(xi)滌(di)液(ye)(ye)(ye)(ye)(ye)(ye)，吸(xi)水紙(zhi)上(shang)(shang)拍(pai)干(gan)，如此重復(fu)洗(xi)(xi)(xi)板(ban)(ban)(ban)5次(ci)（也可用(yong)(yong)(yong)洗(xi)(xi)(xi)板(ban)(ban)(ban)機洗(xi)(xi)(xi)板(ban)(ban)(ban)）。6.每孔(kong)(kong)(kong)(kong)加(jia)(jia)(jia)入(ru)底物(wu)A、B各50μL，37℃避(bi)光孵育(yu)(yu)15min。7.每孔(kong)(kong)(kong)(kong)加(jia)(jia)(jia)入(ru)終(zhong)止液(ye)(ye)(ye)(ye)(ye)(ye)50μL，15min內(nei)，在(zai)450nm波長(chang)處(chu)測(ce)定(ding)各孔(kong)(kong)(kong)(kong)的(de)OD值(zhi)。結果判斷 繪(hui)制標(biao)(biao)(biao)(biao)準(zhun)曲線：在(zai)Excel工作(zuo)(zuo)表中(zhong)，以(yi)標(biao)(biao)(biao)(biao)準(zhun)品(pin)(pin)(pin)濃(nong)(nong)度(du)(du)作(zuo)(zuo)橫坐(zuo)標(biao)(biao)(biao)(biao)，對應OD值(zhi)作(zuo)(zuo)縱(zong)坐(zuo)標(biao)(biao)(biao)(biao)，繪(hui)制出(chu)標(biao)(biao)(biao)(biao)準(zhun)品(pin)(pin)(pin)線性回(hui)(hui)歸曲線，按(an)(an)曲線方(fang)程計(ji)(ji)算(suan)各樣本濃(nong)(nong)度(du)(du)值(zhi)。大(da)鼠白(bai)(bai)血(xue)(xue)(xue)病(bing)(bing)抑制因(yin)(yin)子(zi)(zi)受體(ti)(ti)(ti)(ti)(ti)(ti)(ti)(LIFR)ELISA試(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)試(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)性能1.準(zhun)確(que)(que)性：標(biao)(biao)(biao)(biao)準(zhun)品(pin)(pin)(pin)線性回(hui)(hui)歸與(yu)預期(qi)濃(nong)(nong)度(du)(du)相關系(xi)數R值(zhi)，大(da)于(yu)等(deng)于(yu)0.9900。2.靈敏度(du)(du)：＊低檢(jian)測(ce)濃(nong)(nong)度(du)(du)小于(yu)1.0 U/ml。3.特異性：不(bu)(bu)(bu)與(yu)其(qi)它可溶性結構類(lei)似物(wu)交叉反(fan)(fan)應。4.重復(fu)性：板(ban)(ban)(ban)內(nei)、板(ban)(ban)(ban)間變異系(xi)數均小于(yu)15%。5.貯藏：2-8℃，避(bi)光防潮(chao)保(bao)(bao)存(cun)。6.有(you)效期(qi)：6個月免(mian)(mian)責(ze)聲明(ming)1.試(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)僅供(gong)研究使用(yong)(yong)(yong)，不(bu)(bu)(bu)得用(yong)(yong)(yong)于(yu)臨(lin)床實(shi)(shi)驗(yan)或(huo)人體(ti)(ti)(ti)(ti)(ti)(ti)(ti)實(shi)(shi)驗(yan)，否則所(suo)產生(sheng)的(de)一切后(hou)果，由實(shi)(shi)驗(yan)者(zhe)承(cheng)擔，本公司概不(bu)(bu)(bu)負(fu)責(ze)。2.嚴格(ge)按(an)(an)照說(shuo)(shuo)明(ming)書(shu)(shu)操(cao)(cao)(cao)作(zuo)(zuo)，實(shi)(shi)驗(yan)者(zhe)違反(fan)(fan)說(shuo)(shuo)明(ming)書(shu)(shu)操(cao)(cao)(cao)作(zuo)(zuo)，后(hou)果由實(shi)(shi)驗(yan)者(zhe)承(cheng)擔。 大(da)鼠白(bai)(bai)血(xue)(xue)(xue)病(bing)(bing)抑制因(yin)(yin)子(zi)(zi)受體(ti)(ti)(ti)(ti)(ti)(ti)(ti)(LIFR)ELISA試(shi)(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)
大鼠(shu)鉤(gou)端(duan)螺(luo)旋(xuan)(xuan)(xuan)體(ti)IgG(Lep IgG)ELISA試(shi)(shi)劑(ji)(ji)(ji)(ji)盒(he)(he)(he)大鼠(shu)鉤(gou)端(duan)螺(luo)旋(xuan)(xuan)(xuan)體(ti)IgG(Lep IgG)ELISA試(shi)(shi)劑(ji)(ji)(ji)(ji)盒(he)(he)(he)檢(jian)測(ce)原(yuan)理試(shi)(shi)劑(ji)(ji)(ji)(ji)盒(he)(he)(he)采用(yong)(yong)(yong)雙抗(kang)體(ti)一(yi)(yi)步夾心法酶聯免疫吸(xi)(xi)(xi)附(fu)試(shi)(shi)驗(yan)（ELISA）。往預(yu)先包被活性(xing)(xing)(xing)氧(yang)(yang)簇（ROS）抗(kang)體(ti)的(de)(de)(de)包被微孔(kong)(kong)(kong)中(zhong)，依(yi)(yi)次(ci)(ci)加(jia)(jia)(jia)入標(biao)(biao)(biao)(biao)(biao)本(ben)(ben)(ben)、標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)、HRP標(biao)(biao)(biao)(biao)(biao)記的(de)(de)(de)檢(jian)測(ce)抗(kang)體(ti)，經過溫育并(bing)徹底(di)(di)洗(xi)(xi)(xi)滌(di)(di)(di)。用(yong)(yong)(yong)底(di)(di)物(wu)(wu)(wu)(wu)TMB顯色(se)，TMB在(zai)(zai)過氧(yang)(yang)化(hua)(hua)物(wu)(wu)(wu)(wu)酶的(de)(de)(de)催化(hua)(hua)下轉化(hua)(hua)成藍色(se)，并(bing)在(zai)(zai)酸(suan)的(de)(de)(de)作(zuo)(zuo)用(yong)(yong)(yong)下轉化(hua)(hua)成＊終(zhong)的(de)(de)(de)黃色(se)。顏(yan)色(se)的(de)(de)(de)深(shen)淺(qian)和樣(yang)品(pin)(pin)(pin)中(zhong)的(de)(de)(de)活性(xing)(xing)(xing)氧(yang)(yang)簇（ROS）呈正(zheng)相(xiang)關。用(yong)(yong)(yong)酶標(biao)(biao)(biao)(biao)(biao)儀在(zai)(zai)450nm 波長下測(ce)定吸(xi)(xi)(xi)光度(du)(du)(du)(du)(du)（OD 值(zhi)），計算樣(yang)品(pin)(pin)(pin)活性(xing)(xing)(xing)。大鼠(shu)鉤(gou)端(duan)螺(luo)旋(xuan)(xuan)(xuan)體(ti)IgG(Lep IgG)ELISA試(shi)(shi)劑(ji)(ji)(ji)(ji)盒(he)(he)(he)樣(yang)品(pin)(pin)(pin)收集、處理及(ji)保(bao)(bao)存(cun)(cun)方法1. 血清(qing)：使(shi)用(yong)(yong)(yong)不(bu)含熱(re)原(yuan)和內(nei)(nei)毒素的(de)(de)(de)試(shi)(shi)管，操(cao)作(zuo)(zuo)過程中(zhong)避免任何細(xi)胞(bao)刺(ci)激(ji)，收集血液(ye)(ye)(ye)(ye)后(hou)(hou)，3000轉離(li)心10分鐘將血清(qing)和紅細(xi)胞(bao)迅速(su)小心地(di)分離(li)。2. 血漿：EDTA、檸(ning)檬(meng)酸(suan)鹽(yan)或(huo)(huo)肝素抗(kang)凝。3000轉離(li)心30分鐘取上清(qing)。3. 細(xi)胞(bao)上清(qing)液(ye)(ye)(ye)(ye)：3000轉離(li)心10分鐘去(qu)除顆粒(li)和聚合(he)物(wu)(wu)(wu)(wu)。4. 組織(zhi)勻漿：將組織(zhi)加(jia)(jia)(jia)入適量(liang)生(sheng)理鹽(yan)水搗碎。3000轉離(li)心10分鐘取上清(qing)。5. 保(bao)(bao)存(cun)(cun)：如(ru)(ru)果(guo)樣(yang)本(ben)(ben)(ben)收集后(hou)(hou)不(bu)及(ji)時(shi)檢(jian)測(ce)，請按一(yi)(yi)次(ci)(ci)用(yong)(yong)(yong)量(liang)分裝，凍存(cun)(cun)于(yu)(yu)-20℃，避免反(fan)復(fu)凍融(rong)，在(zai)(zai)室溫下解凍并(bing)確保(bao)(bao)樣(yang)品(pin)(pin)(pin)均勻地(di)充分解凍。大鼠(shu)鉤(gou)端(duan)螺(luo)旋(xuan)(xuan)(xuan)體(ti)IgG(Lep IgG)ELISA試(shi)(shi)劑(ji)(ji)(ji)(ji)盒(he)(he)(he)自(zi)備物(wu)(wu)(wu)(wu)品(pin)(pin)(pin)1.酶標(biao)(biao)(biao)(biao)(biao)儀（450nm）2.高精度(du)(du)(du)(du)(du)加(jia)(jia)(jia)樣(yang)器及(ji)槍頭(tou)：0.5-10uL、2-20uL、20-200uL、200-1000uL3.37℃恒溫箱(xiang)操(cao)作(zuo)(zuo)注意事(shi)項1.試(shi)(shi)劑(ji)(ji)(ji)(ji)盒(he)(he)(he)保(bao)(bao)存(cun)(cun)在(zai)(zai)2-8℃，使(shi)用(yong)(yong)(yong)前(qian)室溫平衡20分鐘。從冰箱(xiang)取出(chu)的(de)(de)(de)濃(nong)(nong)縮洗(xi)(xi)(xi)滌(di)(di)(di)液(ye)(ye)(ye)(ye)會(hui)有結(jie)(jie)晶，這屬于(yu)(yu)正(zheng)常現象，水浴加(jia)(jia)(jia)熱(re)使(shi)結(jie)(jie)晶完全溶解后(hou)(hou)再使(shi)用(yong)(yong)(yong)。2.實驗(yan)中(zhong)不(bu)用(yong)(yong)(yong)的(de)(de)(de)板(ban)(ban)條(tiao)應(ying)(ying)(ying)立即(ji)放回(hui)(hui)(hui)自(zi)封(feng)(feng)(feng)袋(dai)(dai)(dai)中(zhong)，密(mi)封(feng)(feng)(feng)（低溫干(gan)燥）保(bao)(bao)存(cun)(cun)。3.濃(nong)(nong)度(du)(du)(du)(du)(du)為0的(de)(de)(de)S0號(hao)標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)即(ji)可視為陰性(xing)(xing)(xing)對(dui)照(zhao)(zhao)或(huo)(huo)者(zhe)(zhe)空(kong)(kong)白(bai)；按照(zhao)(zhao)說(shuo)(shuo)明(ming)(ming)書操(cao)作(zuo)(zuo)時(shi)樣(yang)本(ben)(ben)(ben)已經稀釋5倍(bei)，＊終(zhong)結(jie)(jie)果(guo)乘(cheng)以5才是(shi)樣(yang)本(ben)(ben)(ben)實際濃(nong)(nong)度(du)(du)(du)(du)(du)。4.嚴格按照(zhao)(zhao)說(shuo)(shuo)明(ming)(ming)書中(zhong)標(biao)(biao)(biao)(biao)(biao)明(ming)(ming)的(de)(de)(de)時(shi)間(jian)、加(jia)(jia)(jia)液(ye)(ye)(ye)(ye)量(liang)及(ji)順序進(jin)行溫育操(cao)作(zuo)(zuo)。5.所有液(ye)(ye)(ye)(ye)體(ti)組分使(shi)用(yong)(yong)(yong)前(qian)充分搖勻。試(shi)(shi)劑(ji)(ji)(ji)(ji)盒(he)(he)(he)組成名稱96孔(kong)(kong)(kong)配置(zhi)(zhi)48孔(kong)(kong)(kong)配置(zhi)(zhi)備注微孔(kong)(kong)(kong)酶標(biao)(biao)(biao)(biao)(biao)板(ban)(ban)12孔(kong)(kong)(kong)×8條(tiao)12孔(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)0.3mL*6管0.3mL*6管無(wu)(wu)(wu)(wu)樣(yang)本(ben)(ben)(ben)稀釋液(ye)(ye)(ye)(ye)6mL3mL無(wu)(wu)(wu)(wu)檢(jian)測(ce)抗(kang)體(ti)-HRP10mL5mL無(wu)(wu)(wu)(wu)20×洗(xi)(xi)(xi)滌(di)(di)(di)緩(huan)沖(chong)液(ye)(ye)(ye)(ye)25mL15mL按說(shuo)(shuo)明(ming)(ming)書進(jin)行稀釋底(di)(di)物(wu)(wu)(wu)(wu)A6mL3mL無(wu)(wu)(wu)(wu)底(di)(di)物(wu)(wu)(wu)(wu)B6mL3mL無(wu)(wu)(wu)(wu)終(zhong)止液(ye)(ye)(ye)(ye)6mL3mL無(wu)(wu)(wu)(wu)封(feng)(feng)(feng)板(ban)(ban)膜2張2張無(wu)(wu)(wu)(wu)說(shuo)(shuo)明(ming)(ming)書1份1份無(wu)(wu)(wu)(wu)自(zi)封(feng)(feng)(feng)袋(dai)(dai)(dai)1個1個無(wu)(wu)(wu)(wu)注：標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)（S0-S5）濃(nong)(nong)度(du)(du)(du)(du)(du)依(yi)(yi)次(ci)(ci)為：0、30、60、120、240、480 U/ml試(shi)(shi)劑(ji)(ji)(ji)(ji)的(de)(de)(de)準(zhun)(zhun)(zhun)(zhun)(zhun)備 20×洗(xi)(xi)(xi)滌(di)(di)(di)緩(huan)沖(chong)液(ye)(ye)(ye)(ye)的(de)(de)(de)稀釋：蒸(zheng)(zheng)餾水按1：20稀釋，即(ji)1份的(de)(de)(de)20×洗(xi)(xi)(xi)滌(di)(di)(di)緩(huan)沖(chong)液(ye)(ye)(ye)(ye)加(jia)(jia)(jia)19份的(de)(de)(de)蒸(zheng)(zheng)餾水。洗(xi)(xi)(xi)板(ban)(ban)方法1.手工洗(xi)(xi)(xi)板(ban)(ban)：甩盡(jin)孔(kong)(kong)(kong)內(nei)(nei)液(ye)(ye)(ye)(ye)體(ti)，每孔(kong)(kong)(kong)加(jia)(jia)(jia)滿洗(xi)(xi)(xi)滌(di)(di)(di)液(ye)(ye)(ye)(ye)，靜置(zhi)(zhi)1min后(hou)(hou)甩盡(jin)孔(kong)(kong)(kong)內(nei)(nei)液(ye)(ye)(ye)(ye)體(ti)，在(zai)(zai)吸(xi)(xi)(xi)水紙上拍(pai)干(gan)，如(ru)(ru)此(ci)洗(xi)(xi)(xi)板(ban)(ban)5次(ci)(ci)。2.自(zi)動洗(xi)(xi)(xi)板(ban)(ban)機：每孔(kong)(kong)(kong)注入洗(xi)(xi)(xi)液(ye)(ye)(ye)(ye)350μL，浸泡1min，洗(xi)(xi)(xi)板(ban)(ban)5次(ci)(ci)。操(cao)作(zuo)(zuo)步驟1.從室溫平衡20min后(hou)(hou)的(de)(de)(de)鋁箔袋(dai)(dai)(dai)中(zhong)取出(chu)所需板(ban)(ban)條(tiao)，剩余板(ban)(ban)條(tiao)用(yong)(yong)(yong)自(zi)封(feng)(feng)(feng)袋(dai)(dai)(dai)密(mi)封(feng)(feng)(feng)放回(hui)(hui)(hui)4℃。2.設置(zhi)(zhi)標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)孔(kong)(kong)(kong)和樣(yang)本(ben)(ben)(ben)孔(kong)(kong)(kong)，標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)孔(kong)(kong)(kong)各加(jia)(jia)(jia)不(bu)同濃(nong)(nong)度(du)(du)(du)(du)(du)的(de)(de)(de)標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)50μL；3.樣(yang)本(ben)(ben)(ben)孔(kong)(kong)(kong)先加(jia)(jia)(jia)待測(ce)樣(yang)本(ben)(ben)(ben)10μL，再加(jia)(jia)(jia)樣(yang)本(ben)(ben)(ben)稀釋液(ye)(ye)(ye)(ye)40μL；空(kong)(kong)白(bai)孔(kong)(kong)(kong)不(bu)加(jia)(jia)(jia)。4.除空(kong)(kong)白(bai)孔(kong)(kong)(kong)外，標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)孔(kong)(kong)(kong)和樣(yang)本(ben)(ben)(ben)孔(kong)(kong)(kong)中(zhong)每孔(kong)(kong)(kong)加(jia)(jia)(jia)入辣根過氧(yang)(yang)化(hua)(hua)物(wu)(wu)(wu)(wu)酶（HRP）標(biao)(biao)(biao)(biao)(biao)記的(de)(de)(de)檢(jian)測(ce)抗(kang)體(ti)100μL，用(yong)(yong)(yong)封(feng)(feng)(feng)板(ban)(ban)膜封(feng)(feng)(feng)住(zhu)反(fan)應(ying)(ying)(ying)孔(kong)(kong)(kong)，37℃水浴鍋或(huo)(huo)恒溫箱(xiang)溫育60min。5.棄去(qu)液(ye)(ye)(ye)(ye)體(ti)，吸(xi)(xi)(xi)水紙上拍(pai)干(gan)，每孔(kong)(kong)(kong)加(jia)(jia)(jia)滿洗(xi)(xi)(xi)滌(di)(di)(di)液(ye)(ye)(ye)(ye)，靜置(zhi)(zhi)1min，甩去(qu)洗(xi)(xi)(xi)滌(di)(di)(di)液(ye)(ye)(ye)(ye)，吸(xi)(xi)(xi)水紙上拍(pai)干(gan)，如(ru)(ru)此(ci)重復(fu)洗(xi)(xi)(xi)板(ban)(ban)5次(ci)(ci)（也可用(yong)(yong)(yong)洗(xi)(xi)(xi)板(ban)(ban)機洗(xi)(xi)(xi)板(ban)(ban)）。6.每孔(kong)(kong)(kong)加(jia)(jia)(jia)入底(di)(di)物(wu)(wu)(wu)(wu)A、B各50μL，37℃避光孵育15min。7.每孔(kong)(kong)(kong)加(jia)(jia)(jia)入終(zhong)止液(ye)(ye)(ye)(ye)50μL，15min內(nei)(nei)，在(zai)(zai)450nm波長處測(ce)定各孔(kong)(kong)(kong)的(de)(de)(de)OD值(zhi)。結(jie)(jie)果(guo)判斷 繪制(zhi)標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)曲線(xian)：在(zai)(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)濃(nong)(nong)度(du)(du)(du)(du)(du)作(zuo)(zuo)橫(heng)坐標(biao)(biao)(biao)(biao)(biao)，對(dui)應(ying)(ying)(ying)OD值(zhi)作(zuo)(zuo)縱坐標(biao)(biao)(biao)(biao)(biao)，繪制(zhi)出(chu)標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)線(xian)性(xing)(xing)(xing)回(hui)(hui)(hui)歸(gui)曲線(xian)，按曲線(xian)方程計算各樣(yang)本(ben)(ben)(ben)濃(nong)(nong)度(du)(du)(du)(du)(du)值(zhi)。大鼠(shu)鉤(gou)端(duan)螺(luo)旋(xuan)(xuan)(xuan)體(ti)IgG(Lep IgG)ELISA試(shi)(shi)劑(ji)(ji)(ji)(ji)盒(he)(he)(he)試(shi)(shi)劑(ji)(ji)(ji)(ji)盒(he)(he)(he)性(xing)(xing)(xing)能1.準(zhun)(zhun)(zhun)(zhun)(zhun)確性(xing)(xing)(xing)：標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)線(xian)性(xing)(xing)(xing)回(hui)(hui)(hui)歸(gui)與(yu)預(yu)期(qi)濃(nong)(nong)度(du)(du)(du)(du)(du)相(xiang)關系數(shu)R值(zhi)，大于(yu)(yu)等于(yu)(yu)0.9900。2.靈敏度(du)(du)(du)(du)(du)：＊低檢(jian)測(ce)濃(nong)(nong)度(du)(du)(du)(du)(du)小于(yu)(yu)1.0 U/ml。3.特異(yi)性(xing)(xing)(xing)：不(bu)與(yu)其它(ta)可溶性(xing)(xing)(xing)結(jie)(jie)構(gou)類似物(wu)(wu)(wu)(wu)交叉反(fan)應(ying)(ying)(ying)。4.重復(fu)性(xing)(xing)(xing)：板(ban)(ban)內(nei)(nei)、板(ban)(ban)間(jian)變異(yi)系數(shu)均小于(yu)(yu)15%。5.貯藏(zang)：2-8℃，避光防潮(chao)保(bao)(bao)存(cun)(cun)。6.有效期(qi)：6個月免責(ze)(ze)聲(sheng)明(ming)(ming)1.試(shi)(shi)劑(ji)(ji)(ji)(ji)盒(he)(he)(he)僅供(gong)研究(jiu)使(shi)用(yong)(yong)(yong)，不(bu)得用(yong)(yong)(yong)于(yu)(yu)臨床(chuang)實驗(yan)或(huo)(huo)人體(ti)實驗(yan)，否則(ze)所產生(sheng)的(de)(de)(de)一(yi)(yi)切(qie)后(hou)(hou)果(guo)，由(you)(you)實驗(yan)者(zhe)(zhe)承擔(dan)，本(ben)(ben)(ben)公(gong)司概不(bu)負責(ze)(ze)。2.嚴格按照(zhao)(zhao)說(shuo)(shuo)明(ming)(ming)書操(cao)作(zuo)(zuo)，實驗(yan)者(zhe)(zhe)違反(fan)說(shuo)(shuo)明(ming)(ming)書操(cao)作(zuo)(zuo)，后(hou)(hou)果(guo)由(you)(you)實驗(yan)者(zhe)(zhe)承擔(dan)。 大鼠(shu)鉤(gou)端(duan)螺(luo)旋(xuan)(xuan)(xuan)體(ti)IgG(Lep IgG)ELISA試(shi)(shi)劑(ji)(ji)(ji)(ji)盒(he)(he)(he)
大(da)鼠(shu)(shu)瘦(shou)(shou)素(su)(LEP)ELISA試(shi)(shi)劑(ji)(ji)(ji)盒(he)(he)(he)大(da)鼠(shu)(shu)瘦(shou)(shou)素(su)(LEP)ELISA試(shi)(shi)劑(ji)(ji)(ji)盒(he)(he)(he)檢測原(yuan)理(li)試(shi)(shi)劑(ji)(ji)(ji)盒(he)(he)(he)采用(yong)(yong)雙抗(kang)(kang)體(ti)(ti)一步夾心(xin)法(fa)酶(mei)(mei)聯免(mian)疫吸(xi)(xi)附試(shi)(shi)驗(yan)(yan)（ELISA）。往預(yu)先包(bao)(bao)被(bei)活(huo)性(xing)(xing)(xing)氧(yang)簇(cu)（ROS）抗(kang)(kang)體(ti)(ti)的(de)(de)(de)(de)(de)包(bao)(bao)被(bei)微(wei)孔(kong)(kong)(kong)(kong)中(zhong)(zhong)(zhong)(zhong)，依次(ci)(ci)(ci)加(jia)入(ru)(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)(pin)、HRP標(biao)(biao)(biao)(biao)(biao)(biao)(biao)記的(de)(de)(de)(de)(de)檢測抗(kang)(kang)體(ti)(ti)，經(jing)過溫(wen)育(yu)并(bing)(bing)徹底(di)洗(xi)(xi)(xi)(xi)滌。用(yong)(yong)底(di)物(wu)TMB顯(xian)色(se)(se)，TMB在(zai)過氧(yang)化物(wu)酶(mei)(mei)的(de)(de)(de)(de)(de)催化下(xia)(xia)(xia)轉化成(cheng)藍(lan)色(se)(se)，并(bing)(bing)在(zai)酸(suan)的(de)(de)(de)(de)(de)作(zuo)(zuo)(zuo)用(yong)(yong)下(xia)(xia)(xia)轉化成(cheng)＊終的(de)(de)(de)(de)(de)黃色(se)(se)。顏色(se)(se)的(de)(de)(de)(de)(de)深淺和(he)(he)樣(yang)(yang)品(pin)(pin)(pin)中(zhong)(zhong)(zhong)(zhong)的(de)(de)(de)(de)(de)活(huo)性(xing)(xing)(xing)氧(yang)簇(cu)（ROS）呈正相關。用(yong)(yong)酶(mei)(mei)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)儀(yi)在(zai)450nm 波(bo)長下(xia)(xia)(xia)測定(ding)吸(xi)(xi)光(guang)度(du)(du)(du)(du)（OD 值(zhi)），計(ji)算樣(yang)(yang)品(pin)(pin)(pin)活(huo)性(xing)(xing)(xing)。大(da)鼠(shu)(shu)瘦(shou)(shou)素(su)(LEP)ELISA試(shi)(shi)劑(ji)(ji)(ji)盒(he)(he)(he)樣(yang)(yang)品(pin)(pin)(pin)收集、處理(li)及(ji)(ji)保(bao)存(cun)(cun)方法(fa)1. 血清(qing)(qing)：使(shi)(shi)用(yong)(yong)不(bu)(bu)含熱原(yuan)和(he)(he)內(nei)毒(du)素(su)的(de)(de)(de)(de)(de)試(shi)(shi)管(guan)，操(cao)(cao)(cao)作(zuo)(zuo)(zuo)過程中(zhong)(zhong)(zhong)(zhong)避(bi)免(mian)任何細胞刺激，收集血液(ye)(ye)(ye)(ye)(ye)(ye)后，3000轉離心(xin)10分(fen)(fen)鐘(zhong)將血清(qing)(qing)和(he)(he)紅(hong)細胞迅(xun)速小心(xin)地分(fen)(fen)離。2. 血漿：EDTA、檸(ning)檬酸(suan)鹽(yan)或肝(gan)素(su)抗(kang)(kang)凝(ning)。3000轉離心(xin)30分(fen)(fen)鐘(zhong)取(qu)(qu)上(shang)清(qing)(qing)。3. 細胞上(shang)清(qing)(qing)液(ye)(ye)(ye)(ye)(ye)(ye)：3000轉離心(xin)10分(fen)(fen)鐘(zhong)去除(chu)(chu)顆粒和(he)(he)聚合物(wu)。4. 組(zu)織勻漿：將組(zu)織加(jia)入(ru)(ru)適(shi)量生(sheng)理(li)鹽(yan)水(shui)搗碎(sui)。3000轉離心(xin)10分(fen)(fen)鐘(zhong)取(qu)(qu)上(shang)清(qing)(qing)。5. 保(bao)存(cun)(cun)：如(ru)果(guo)樣(yang)(yang)本(ben)(ben)收集后不(bu)(bu)及(ji)(ji)時檢測，請按一次(ci)(ci)(ci)用(yong)(yong)量分(fen)(fen)裝，凍存(cun)(cun)于-20℃，避(bi)免(mian)反(fan)復(fu)凍融，在(zai)室溫(wen)下(xia)(xia)(xia)解凍并(bing)(bing)確(que)保(bao)樣(yang)(yang)品(pin)(pin)(pin)均勻地充(chong)分(fen)(fen)解凍。大(da)鼠(shu)(shu)瘦(shou)(shou)素(su)(LEP)ELISA試(shi)(shi)劑(ji)(ji)(ji)盒(he)(he)(he)自(zi)備(bei)(bei)物(wu)品(pin)(pin)(pin)1.酶(mei)(mei)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)儀(yi)（450nm）2.高精(jing)度(du)(du)(du)(du)加(jia)樣(yang)(yang)器及(ji)(ji)槍頭：0.5-10uL、2-20uL、20-200uL、200-1000uL3.37℃恒溫(wen)箱操(cao)(cao)(cao)作(zuo)(zuo)(zuo)注(zhu)意事(shi)項1.試(shi)(shi)劑(ji)(ji)(ji)盒(he)(he)(he)保(bao)存(cun)(cun)在(zai)2-8℃，使(shi)(shi)用(yong)(yong)前室溫(wen)平衡(heng)20分(fen)(fen)鐘(zhong)。從(cong)冰(bing)箱取(qu)(qu)出的(de)(de)(de)(de)(de)濃(nong)縮(suo)洗(xi)(xi)(xi)(xi)滌液(ye)(ye)(ye)(ye)(ye)(ye)會有(you)結晶，這(zhe)屬于正常現象，水(shui)浴(yu)加(jia)熱使(shi)(shi)結晶完全溶解后再(zai)使(shi)(shi)用(yong)(yong)。2.實驗(yan)(yan)中(zhong)(zhong)(zhong)(zhong)不(bu)(bu)用(yong)(yong)的(de)(de)(de)(de)(de)板(ban)(ban)條應(ying)立即(ji)放(fang)回自(zi)封(feng)袋(dai)中(zhong)(zhong)(zhong)(zhong)，密封(feng)（低溫(wen)干燥）保(bao)存(cun)(cun)。3.濃(nong)度(du)(du)(du)(du)為(wei)0的(de)(de)(de)(de)(de)S0號標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)即(ji)可(ke)視為(wei)陰(yin)性(xing)(xing)(xing)對(dui)照(zhao)(zhao)或者空白；按照(zhao)(zhao)說(shuo)明(ming)(ming)書(shu)(shu)操(cao)(cao)(cao)作(zuo)(zuo)(zuo)時樣(yang)(yang)本(ben)(ben)已經(jing)稀釋(shi)(shi)5倍，＊終結果(guo)乘以5才是樣(yang)(yang)本(ben)(ben)實際(ji)濃(nong)度(du)(du)(du)(du)。4.嚴格按照(zhao)(zhao)說(shuo)明(ming)(ming)書(shu)(shu)中(zhong)(zhong)(zhong)(zhong)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)明(ming)(ming)的(de)(de)(de)(de)(de)時間、加(jia)液(ye)(ye)(ye)(ye)(ye)(ye)量及(ji)(ji)順序進行(xing)溫(wen)育(yu)操(cao)(cao)(cao)作(zuo)(zuo)(zuo)。5.所(suo)(suo)有(you)液(ye)(ye)(ye)(ye)(ye)(ye)體(ti)(ti)組(zu)分(fen)(fen)使(shi)(shi)用(yong)(yong)前充(chong)分(fen)(fen)搖(yao)勻。試(shi)(shi)劑(ji)(ji)(ji)盒(he)(he)(he)組(zu)成(cheng)名稱96孔(kong)(kong)(kong)(kong)配置(zhi)(zhi)48孔(kong)(kong)(kong)(kong)配置(zhi)(zhi)備(bei)(bei)注(zhu)微(wei)孔(kong)(kong)(kong)(kong)酶(mei)(mei)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)板(ban)(ban)12孔(kong)(kong)(kong)(kong)×8條12孔(kong)(kong)(kong)(kong)×4條無(wu)(wu)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)0.3mL*6管(guan)0.3mL*6管(guan)無(wu)(wu)樣(yang)(yang)本(ben)(ben)稀釋(shi)(shi)液(ye)(ye)(ye)(ye)(ye)(ye)6mL3mL無(wu)(wu)檢測抗(kang)(kang)體(ti)(ti)-HRP10mL5mL無(wu)(wu)20×洗(xi)(xi)(xi)(xi)滌緩沖液(ye)(ye)(ye)(ye)(ye)(ye)25mL15mL按說(shuo)明(ming)(ming)書(shu)(shu)進行(xing)稀釋(shi)(shi)底(di)物(wu)A6mL3mL無(wu)(wu)底(di)物(wu)B6mL3mL無(wu)(wu)終止液(ye)(ye)(ye)(ye)(ye)(ye)6mL3mL無(wu)(wu)封(feng)板(ban)(ban)膜2張(zhang)2張(zhang)無(wu)(wu)說(shuo)明(ming)(ming)書(shu)(shu)1份1份無(wu)(wu)自(zi)封(feng)袋(dai)1個1個無(wu)(wu)注(zhu)：標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)（S0-S5）濃(nong)度(du)(du)(du)(du)依次(ci)(ci)(ci)為(wei)：0、30、60、120、240、480 U/ml試(shi)(shi)劑(ji)(ji)(ji)的(de)(de)(de)(de)(de)準(zhun)(zhun)(zhun)(zhun)備(bei)(bei) 20×洗(xi)(xi)(xi)(xi)滌緩沖液(ye)(ye)(ye)(ye)(ye)(ye)的(de)(de)(de)(de)(de)稀釋(shi)(shi)：蒸餾(liu)水(shui)按1：20稀釋(shi)(shi)，即(ji)1份的(de)(de)(de)(de)(de)20×洗(xi)(xi)(xi)(xi)滌緩沖液(ye)(ye)(ye)(ye)(ye)(ye)加(jia)19份的(de)(de)(de)(de)(de)蒸餾(liu)水(shui)。洗(xi)(xi)(xi)(xi)板(ban)(ban)方法(fa)1.手工(gong)洗(xi)(xi)(xi)(xi)板(ban)(ban)：甩(shuai)盡(jin)(jin)孔(kong)(kong)(kong)(kong)內(nei)液(ye)(ye)(ye)(ye)(ye)(ye)體(ti)(ti)，每(mei)(mei)孔(kong)(kong)(kong)(kong)加(jia)滿洗(xi)(xi)(xi)(xi)滌液(ye)(ye)(ye)(ye)(ye)(ye)，靜(jing)置(zhi)(zhi)1min后甩(shuai)盡(jin)(jin)孔(kong)(kong)(kong)(kong)內(nei)液(ye)(ye)(ye)(ye)(ye)(ye)體(ti)(ti)，在(zai)吸(xi)(xi)水(shui)紙(zhi)上(shang)拍干，如(ru)此(ci)洗(xi)(xi)(xi)(xi)板(ban)(ban)5次(ci)(ci)(ci)。2.自(zi)動洗(xi)(xi)(xi)(xi)板(ban)(ban)機：每(mei)(mei)孔(kong)(kong)(kong)(kong)注(zhu)入(ru)(ru)洗(xi)(xi)(xi)(xi)液(ye)(ye)(ye)(ye)(ye)(ye)350μL，浸泡1min，洗(xi)(xi)(xi)(xi)板(ban)(ban)5次(ci)(ci)(ci)。操(cao)(cao)(cao)作(zuo)(zuo)(zuo)步驟1.從(cong)室溫(wen)平衡(heng)20min后的(de)(de)(de)(de)(de)鋁箔(bo)袋(dai)中(zhong)(zhong)(zhong)(zhong)取(qu)(qu)出所(suo)(suo)需板(ban)(ban)條，剩余(yu)板(ban)(ban)條用(yong)(yong)自(zi)封(feng)袋(dai)密封(feng)放(fang)回4℃。2.設(she)置(zhi)(zhi)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)孔(kong)(kong)(kong)(kong)和(he)(he)樣(yang)(yang)本(ben)(ben)孔(kong)(kong)(kong)(kong)，標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)孔(kong)(kong)(kong)(kong)各加(jia)不(bu)(bu)同濃(nong)度(du)(du)(du)(du)的(de)(de)(de)(de)(de)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)50μL；3.樣(yang)(yang)本(ben)(ben)孔(kong)(kong)(kong)(kong)先加(jia)待測樣(yang)(yang)本(ben)(ben)10μL，再(zai)加(jia)樣(yang)(yang)本(ben)(ben)稀釋(shi)(shi)液(ye)(ye)(ye)(ye)(ye)(ye)40μL；空白孔(kong)(kong)(kong)(kong)不(bu)(bu)加(jia)。4.除(chu)(chu)空白孔(kong)(kong)(kong)(kong)外，標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)孔(kong)(kong)(kong)(kong)和(he)(he)樣(yang)(yang)本(ben)(ben)孔(kong)(kong)(kong)(kong)中(zhong)(zhong)(zhong)(zhong)每(mei)(mei)孔(kong)(kong)(kong)(kong)加(jia)入(ru)(ru)辣根(gen)過氧(yang)化物(wu)酶(mei)(mei)（HRP）標(biao)(biao)(biao)(biao)(biao)(biao)(biao)記的(de)(de)(de)(de)(de)檢測抗(kang)(kang)體(ti)(ti)100μL，用(yong)(yong)封(feng)板(ban)(ban)膜封(feng)住反(fan)應(ying)孔(kong)(kong)(kong)(kong)，37℃水(shui)浴(yu)鍋或恒溫(wen)箱溫(wen)育(yu)60min。5.棄去液(ye)(ye)(ye)(ye)(ye)(ye)體(ti)(ti)，吸(xi)(xi)水(shui)紙(zhi)上(shang)拍干，每(mei)(mei)孔(kong)(kong)(kong)(kong)加(jia)滿洗(xi)(xi)(xi)(xi)滌液(ye)(ye)(ye)(ye)(ye)(ye)，靜(jing)置(zhi)(zhi)1min，甩(shuai)去洗(xi)(xi)(xi)(xi)滌液(ye)(ye)(ye)(ye)(ye)(ye)，吸(xi)(xi)水(shui)紙(zhi)上(shang)拍干，如(ru)此(ci)重(zhong)(zhong)復(fu)洗(xi)(xi)(xi)(xi)板(ban)(ban)5次(ci)(ci)(ci)（也可(ke)用(yong)(yong)洗(xi)(xi)(xi)(xi)板(ban)(ban)機洗(xi)(xi)(xi)(xi)板(ban)(ban)）。6.每(mei)(mei)孔(kong)(kong)(kong)(kong)加(jia)入(ru)(ru)底(di)物(wu)A、B各50μL，37℃避(bi)光(guang)孵育(yu)15min。7.每(mei)(mei)孔(kong)(kong)(kong)(kong)加(jia)入(ru)(ru)終止液(ye)(ye)(ye)(ye)(ye)(ye)50μL，15min內(nei)，在(zai)450nm波(bo)長處測定(ding)各孔(kong)(kong)(kong)(kong)的(de)(de)(de)(de)(de)OD值(zhi)。結果(guo)判斷 繪制標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)曲(qu)線(xian)：在(zai)Excel工(gong)作(zuo)(zuo)(zuo)表中(zhong)(zhong)(zhong)(zhong)，以標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)濃(nong)度(du)(du)(du)(du)作(zuo)(zuo)(zuo)橫坐標(biao)(biao)(biao)(biao)(biao)(biao)(biao)，對(dui)應(ying)OD值(zhi)作(zuo)(zuo)(zuo)縱坐標(biao)(biao)(biao)(biao)(biao)(biao)(biao)，繪制出標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)線(xian)性(xing)(xing)(xing)回歸曲(qu)線(xian)，按曲(qu)線(xian)方程計(ji)算各樣(yang)(yang)本(ben)(ben)濃(nong)度(du)(du)(du)(du)值(zhi)。大(da)鼠(shu)(shu)瘦(shou)(shou)素(su)(LEP)ELISA試(shi)(shi)劑(ji)(ji)(ji)盒(he)(he)(he)試(shi)(shi)劑(ji)(ji)(ji)盒(he)(he)(he)性(xing)(xing)(xing)能1.準(zhun)(zhun)(zhun)(zhun)確(que)性(xing)(xing)(xing)：標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)線(xian)性(xing)(xing)(xing)回歸與(yu)預(yu)期(qi)濃(nong)度(du)(du)(du)(du)相關系數(shu)R值(zhi)，大(da)于等(deng)于0.9900。2.靈(ling)敏度(du)(du)(du)(du)：＊低檢測濃(nong)度(du)(du)(du)(du)小于1.0 U/ml。3.特異(yi)性(xing)(xing)(xing)：不(bu)(bu)與(yu)其它(ta)可(ke)溶性(xing)(xing)(xing)結構(gou)類似物(wu)交叉(cha)反(fan)應(ying)。4.重(zhong)(zhong)復(fu)性(xing)(xing)(xing)：板(ban)(ban)內(nei)、板(ban)(ban)間變異(yi)系數(shu)均小于15%。5.貯藏：2-8℃，避(bi)光(guang)防潮保(bao)存(cun)(cun)。6.有(you)效期(qi)：6個月免(mian)責(ze)聲明(ming)(ming)1.試(shi)(shi)劑(ji)(ji)(ji)盒(he)(he)(he)僅(jin)供(gong)研究(jiu)使(shi)(shi)用(yong)(yong)，不(bu)(bu)得用(yong)(yong)于臨床實驗(yan)(yan)或人體(ti)(ti)實驗(yan)(yan)，否則所(suo)(suo)產生(sheng)的(de)(de)(de)(de)(de)一切后果(guo)，由實驗(yan)(yan)者承擔(dan)(dan)，本(ben)(ben)公司概不(bu)(bu)負責(ze)。2.嚴格按照(zhao)(zhao)說(shuo)明(ming)(ming)書(shu)(shu)操(cao)(cao)(cao)作(zuo)(zuo)(zuo)，實驗(yan)(yan)者違反(fan)說(shuo)明(ming)(ming)書(shu)(shu)操(cao)(cao)(cao)作(zuo)(zuo)(zuo)，后果(guo)由實驗(yan)(yan)者承擔(dan)(dan)。 大(da)鼠(shu)(shu)瘦(shou)(shou)素(su)(LEP)ELISA試(shi)(shi)劑(ji)(ji)(ji)盒(he)(he)(he)
大鼠低密(mi)(mi)(mi)(mi)度(du)(du)(du)(du)(du)脂(zhi)(zhi)(zhi)蛋(dan)白(bai)(bai)免(mian)(mian)疫復(fu)(fu)合(he)物(wu)(wu)(wu)(wu)(wu)(LDL-IC) ELISA 試(shi)(shi)(shi)劑(ji)(ji)盒(he) 大鼠低密(mi)(mi)(mi)(mi)度(du)(du)(du)(du)(du)脂(zhi)(zhi)(zhi)蛋(dan)白(bai)(bai)免(mian)(mian)疫復(fu)(fu)合(he)物(wu)(wu)(wu)(wu)(wu)(LDL-IC) ELISA 試(shi)(shi)(shi)劑(ji)(ji)盒(he) 檢(jian)測(ce)原理試(shi)(shi)(shi)劑(ji)(ji)盒(he)采用(yong)(yong)(yong)雙抗(kang)體(ti)(ti)(ti)一(yi)步(bu)夾心(xin)(xin)法(fa)(fa)酶聯免(mian)(mian)疫吸(xi)附試(shi)(shi)(shi)驗（ELISA）。往預先(xian)包(bao)被(bei)活(huo)(huo)性(xing)(xing)(xing)氧(yang)(yang)簇（ROS）抗(kang)體(ti)(ti)(ti)的(de)(de)(de)(de)(de)(de)(de)包(bao)被(bei)微孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)(zhong)(zhong)，依次(ci)(ci)加(jia)(jia)(jia)(jia)(jia)入(ru)標(biao)(biao)(biao)本(ben)(ben)(ben)、標(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)、HRP標(biao)(biao)(biao)記的(de)(de)(de)(de)(de)(de)(de)檢(jian)測(ce)抗(kang)體(ti)(ti)(ti)，經過(guo)(guo)溫(wen)(wen)(wen)育(yu)并(bing)徹(che)底洗(xi)(xi)(xi)(xi)滌(di)。用(yong)(yong)(yong)底物(wu)(wu)(wu)(wu)(wu)TMB顯(xian)色，TMB在(zai)(zai)(zai)過(guo)(guo)氧(yang)(yang)化物(wu)(wu)(wu)(wu)(wu)酶的(de)(de)(de)(de)(de)(de)(de)催化下轉化成藍色，并(bing)在(zai)(zai)(zai)酸(suan)的(de)(de)(de)(de)(de)(de)(de)作(zuo)(zuo)(zuo)用(yong)(yong)(yong)下轉化成＊終的(de)(de)(de)(de)(de)(de)(de)黃色。顏(yan)色的(de)(de)(de)(de)(de)(de)(de)深淺和(he)樣(yang)(yang)(yang)(yang)(yang)品(pin)(pin)(pin)中(zhong)(zhong)(zhong)(zhong)的(de)(de)(de)(de)(de)(de)(de)活(huo)(huo)性(xing)(xing)(xing)氧(yang)(yang)簇（ROS）呈正相關。用(yong)(yong)(yong)酶標(biao)(biao)(biao)儀(yi)在(zai)(zai)(zai)450nm 波長(chang)下測(ce)定吸(xi)光度(du)(du)(du)(du)(du)（OD 值(zhi)），計算樣(yang)(yang)(yang)(yang)(yang)品(pin)(pin)(pin)活(huo)(huo)性(xing)(xing)(xing)。大鼠低密(mi)(mi)(mi)(mi)度(du)(du)(du)(du)(du)脂(zhi)(zhi)(zhi)蛋(dan)白(bai)(bai)免(mian)(mian)疫復(fu)(fu)合(he)物(wu)(wu)(wu)(wu)(wu)(LDL-IC) ELISA 試(shi)(shi)(shi)劑(ji)(ji)盒(he) 樣(yang)(yang)(yang)(yang)(yang)品(pin)(pin)(pin)收(shou)集(ji)、處理及保存方(fang)法(fa)(fa)1. 血(xue)清(qing)(qing)：使(shi)用(yong)(yong)(yong)不(bu)(bu)含熱原和(he)內毒素的(de)(de)(de)(de)(de)(de)(de)試(shi)(shi)(shi)管(guan)(guan)，操(cao)(cao)(cao)作(zuo)(zuo)(zuo)過(guo)(guo)程中(zhong)(zhong)(zhong)(zhong)避免(mian)(mian)任何細胞刺激，收(shou)集(ji)血(xue)液(ye)(ye)(ye)(ye)后(hou)，3000轉離(li)心(xin)(xin)10分(fen)(fen)(fen)鐘(zhong)(zhong)(zhong)將血(xue)清(qing)(qing)和(he)紅細胞迅(xun)速小(xiao)心(xin)(xin)地分(fen)(fen)(fen)離(li)。2. 血(xue)漿：EDTA、檸檬(meng)酸(suan)鹽(yan)或肝素抗(kang)凝。3000轉離(li)心(xin)(xin)30分(fen)(fen)(fen)鐘(zhong)(zhong)(zhong)取(qu)上(shang)(shang)清(qing)(qing)。3. 細胞上(shang)(shang)清(qing)(qing)液(ye)(ye)(ye)(ye)：3000轉離(li)心(xin)(xin)10分(fen)(fen)(fen)鐘(zhong)(zhong)(zhong)去除(chu)顆(ke)粒和(he)聚合(he)物(wu)(wu)(wu)(wu)(wu)。4. 組織勻漿：將組織加(jia)(jia)(jia)(jia)(jia)入(ru)適量生理鹽(yan)水(shui)搗碎。3000轉離(li)心(xin)(xin)10分(fen)(fen)(fen)鐘(zhong)(zhong)(zhong)取(qu)上(shang)(shang)清(qing)(qing)。5. 保存：如(ru)果樣(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)收(shou)集(ji)后(hou)不(bu)(bu)及時檢(jian)測(ce)，請(qing)按(an)一(yi)次(ci)(ci)用(yong)(yong)(yong)量分(fen)(fen)(fen)裝(zhuang)，凍(dong)存于(yu)-20℃，避免(mian)(mian)反復(fu)(fu)凍(dong)融，在(zai)(zai)(zai)室溫(wen)(wen)(wen)下解(jie)凍(dong)并(bing)確保樣(yang)(yang)(yang)(yang)(yang)品(pin)(pin)(pin)均(jun)(jun)勻地充(chong)分(fen)(fen)(fen)解(jie)凍(dong)。大鼠低密(mi)(mi)(mi)(mi)度(du)(du)(du)(du)(du)脂(zhi)(zhi)(zhi)蛋(dan)白(bai)(bai)免(mian)(mian)疫復(fu)(fu)合(he)物(wu)(wu)(wu)(wu)(wu)(LDL-IC) ELISA 試(shi)(shi)(shi)劑(ji)(ji)盒(he) 自備(bei)物(wu)(wu)(wu)(wu)(wu)品(pin)(pin)(pin)1.酶標(biao)(biao)(biao)儀(yi)（450nm）2.高精度(du)(du)(du)(du)(du)加(jia)(jia)(jia)(jia)(jia)樣(yang)(yang)(yang)(yang)(yang)器及槍頭：0.5-10uL、2-20uL、20-200uL、200-1000uL3.37℃恒溫(wen)(wen)(wen)箱(xiang)(xiang)操(cao)(cao)(cao)作(zuo)(zuo)(zuo)注意(yi)事(shi)項(xiang)1.試(shi)(shi)(shi)劑(ji)(ji)盒(he)保存在(zai)(zai)(zai)2-8℃，使(shi)用(yong)(yong)(yong)前室溫(wen)(wen)(wen)平(ping)衡20分(fen)(fen)(fen)鐘(zhong)(zhong)(zhong)。從冰箱(xiang)(xiang)取(qu)出的(de)(de)(de)(de)(de)(de)(de)濃(nong)(nong)縮洗(xi)(xi)(xi)(xi)滌(di)液(ye)(ye)(ye)(ye)會(hui)有結晶，這(zhe)屬(shu)于(yu)正常現象，水(shui)浴(yu)加(jia)(jia)(jia)(jia)(jia)熱使(shi)結晶完全溶解(jie)后(hou)再使(shi)用(yong)(yong)(yong)。2.實(shi)(shi)驗中(zhong)(zhong)(zhong)(zhong)不(bu)(bu)用(yong)(yong)(yong)的(de)(de)(de)(de)(de)(de)(de)板(ban)(ban)(ban)條應(ying)立即(ji)放(fang)回(hui)(hui)自封袋中(zhong)(zhong)(zhong)(zhong)，密(mi)(mi)(mi)(mi)封（低溫(wen)(wen)(wen)干燥(zao)）保存。3.濃(nong)(nong)度(du)(du)(du)(du)(du)為0的(de)(de)(de)(de)(de)(de)(de)S0號標(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)即(ji)可(ke)(ke)視為陰性(xing)(xing)(xing)對照(zhao)(zhao)或者(zhe)空(kong)白(bai)(bai)；按(an)照(zhao)(zhao)說(shuo)(shuo)明(ming)(ming)(ming)書(shu)操(cao)(cao)(cao)作(zuo)(zuo)(zuo)時樣(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)已經稀釋5倍(bei)，＊終結果乘以(yi)5才(cai)是樣(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)實(shi)(shi)際(ji)濃(nong)(nong)度(du)(du)(du)(du)(du)。4.嚴格按(an)照(zhao)(zhao)說(shuo)(shuo)明(ming)(ming)(ming)書(shu)中(zhong)(zhong)(zhong)(zhong)標(biao)(biao)(biao)明(ming)(ming)(ming)的(de)(de)(de)(de)(de)(de)(de)時間(jian)、加(jia)(jia)(jia)(jia)(jia)液(ye)(ye)(ye)(ye)量及順序(xu)進(jin)(jin)行溫(wen)(wen)(wen)育(yu)操(cao)(cao)(cao)作(zuo)(zuo)(zuo)。5.所(suo)有液(ye)(ye)(ye)(ye)體(ti)(ti)(ti)組分(fen)(fen)(fen)使(shi)用(yong)(yong)(yong)前充(chong)分(fen)(fen)(fen)搖勻。試(shi)(shi)(shi)劑(ji)(ji)盒(he)組成名稱96孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)(kong)配置(zhi)48孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)(kong)配置(zhi)備(bei)注微孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)(kong)酶標(biao)(biao)(biao)板(ban)(ban)(ban)12孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)(kong)×8條12孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)(kong)×4條無(wu)(wu)(wu)標(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)0.3mL*6管(guan)(guan)0.3mL*6管(guan)(guan)無(wu)(wu)(wu)樣(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)稀釋液(ye)(ye)(ye)(ye)6mL3mL無(wu)(wu)(wu)檢(jian)測(ce)抗(kang)體(ti)(ti)(ti)-HRP10mL5mL無(wu)(wu)(wu)20×洗(xi)(xi)(xi)(xi)滌(di)緩沖液(ye)(ye)(ye)(ye)25mL15mL按(an)說(shuo)(shuo)明(ming)(ming)(ming)書(shu)進(jin)(jin)行稀釋底物(wu)(wu)(wu)(wu)(wu)A6mL3mL無(wu)(wu)(wu)底物(wu)(wu)(wu)(wu)(wu)B6mL3mL無(wu)(wu)(wu)終止(zhi)液(ye)(ye)(ye)(ye)6mL3mL無(wu)(wu)(wu)封板(ban)(ban)(ban)膜2張2張無(wu)(wu)(wu)說(shuo)(shuo)明(ming)(ming)(ming)書(shu)1份(fen)1份(fen)無(wu)(wu)(wu)自封袋1個1個無(wu)(wu)(wu)注：標(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)（S0-S5）濃(nong)(nong)度(du)(du)(du)(du)(du)依次(ci)(ci)為：0、30、60、120、240、480 U/ml試(shi)(shi)(shi)劑(ji)(ji)的(de)(de)(de)(de)(de)(de)(de)準(zhun)(zhun)(zhun)(zhun)備(bei) 20×洗(xi)(xi)(xi)(xi)滌(di)緩沖液(ye)(ye)(ye)(ye)的(de)(de)(de)(de)(de)(de)(de)稀釋：蒸餾水(shui)按(an)1：20稀釋，即(ji)1份(fen)的(de)(de)(de)(de)(de)(de)(de)20×洗(xi)(xi)(xi)(xi)滌(di)緩沖液(ye)(ye)(ye)(ye)加(jia)(jia)(jia)(jia)(jia)19份(fen)的(de)(de)(de)(de)(de)(de)(de)蒸餾水(shui)。洗(xi)(xi)(xi)(xi)板(ban)(ban)(ban)方(fang)法(fa)(fa)1.手工洗(xi)(xi)(xi)(xi)板(ban)(ban)(ban)：甩盡孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)(kong)內液(ye)(ye)(ye)(ye)體(ti)(ti)(ti)，每孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)(jia)(jia)(jia)滿洗(xi)(xi)(xi)(xi)滌(di)液(ye)(ye)(ye)(ye)，靜置(zhi)1min后(hou)甩盡孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)(kong)內液(ye)(ye)(ye)(ye)體(ti)(ti)(ti)，在(zai)(zai)(zai)吸(xi)水(shui)紙上(shang)(shang)拍(pai)干，如(ru)此(ci)洗(xi)(xi)(xi)(xi)板(ban)(ban)(ban)5次(ci)(ci)。2.自動(dong)洗(xi)(xi)(xi)(xi)板(ban)(ban)(ban)機：每孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)(kong)注入(ru)洗(xi)(xi)(xi)(xi)液(ye)(ye)(ye)(ye)350μL，浸泡(pao)1min，洗(xi)(xi)(xi)(xi)板(ban)(ban)(ban)5次(ci)(ci)。操(cao)(cao)(cao)作(zuo)(zuo)(zuo)步(bu)驟1.從室溫(wen)(wen)(wen)平(ping)衡20min后(hou)的(de)(de)(de)(de)(de)(de)(de)鋁(lv)箔袋中(zhong)(zhong)(zhong)(zhong)取(qu)出所(suo)需板(ban)(ban)(ban)條，剩余(yu)板(ban)(ban)(ban)條用(yong)(yong)(yong)自封袋密(mi)(mi)(mi)(mi)封放(fang)回(hui)(hui)4℃。2.設置(zhi)標(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)(kong)和(he)樣(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)(kong)，標(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)(kong)各(ge)(ge)加(jia)(jia)(jia)(jia)(jia)不(bu)(bu)同濃(nong)(nong)度(du)(du)(du)(du)(du)的(de)(de)(de)(de)(de)(de)(de)標(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)50μL；3.樣(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)(kong)先(xian)加(jia)(jia)(jia)(jia)(jia)待(dai)測(ce)樣(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)10μL，再加(jia)(jia)(jia)(jia)(jia)樣(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)稀釋液(ye)(ye)(ye)(ye)40μL；空(kong)白(bai)(bai)孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)(kong)不(bu)(bu)加(jia)(jia)(jia)(jia)(jia)。4.除(chu)空(kong)白(bai)(bai)孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)(kong)外，標(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)(kong)和(he)樣(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)(zhong)(zhong)每孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)(jia)(jia)(jia)入(ru)辣根過(guo)(guo)氧(yang)(yang)化物(wu)(wu)(wu)(wu)(wu)酶（HRP）標(biao)(biao)(biao)記的(de)(de)(de)(de)(de)(de)(de)檢(jian)測(ce)抗(kang)體(ti)(ti)(ti)100μL，用(yong)(yong)(yong)封板(ban)(ban)(ban)膜封住反應(ying)孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)(kong)，37℃水(shui)浴(yu)鍋或恒溫(wen)(wen)(wen)箱(xiang)(xiang)溫(wen)(wen)(wen)育(yu)60min。5.棄去液(ye)(ye)(ye)(ye)體(ti)(ti)(ti)，吸(xi)水(shui)紙上(shang)(shang)拍(pai)干，每孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)(jia)(jia)(jia)滿洗(xi)(xi)(xi)(xi)滌(di)液(ye)(ye)(ye)(ye)，靜置(zhi)1min，甩去洗(xi)(xi)(xi)(xi)滌(di)液(ye)(ye)(ye)(ye)，吸(xi)水(shui)紙上(shang)(shang)拍(pai)干，如(ru)此(ci)重(zhong)復(fu)(fu)洗(xi)(xi)(xi)(xi)板(ban)(ban)(ban)5次(ci)(ci)（也可(ke)(ke)用(yong)(yong)(yong)洗(xi)(xi)(xi)(xi)板(ban)(ban)(ban)機洗(xi)(xi)(xi)(xi)板(ban)(ban)(ban)）。6.每孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)(jia)(jia)(jia)入(ru)底物(wu)(wu)(wu)(wu)(wu)A、B各(ge)(ge)50μL，37℃避光孵育(yu)15min。7.每孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)(jia)(jia)(jia)入(ru)終止(zhi)液(ye)(ye)(ye)(ye)50μL，15min內，在(zai)(zai)(zai)450nm波長(chang)處測(ce)定各(ge)(ge)孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)(kong)的(de)(de)(de)(de)(de)(de)(de)OD值(zhi)。結果判斷 繪制(zhi)標(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)曲線(xian)：在(zai)(zai)(zai)Excel工作(zuo)(zuo)(zuo)表中(zhong)(zhong)(zhong)(zhong)，以(yi)標(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)濃(nong)(nong)度(du)(du)(du)(du)(du)作(zuo)(zuo)(zuo)橫坐(zuo)標(biao)(biao)(biao)，對應(ying)OD值(zhi)作(zuo)(zuo)(zuo)縱坐(zuo)標(biao)(biao)(biao)，繪制(zhi)出標(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)線(xian)性(xing)(xing)(xing)回(hui)(hui)歸(gui)曲線(xian)，按(an)曲線(xian)方(fang)程計算各(ge)(ge)樣(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)濃(nong)(nong)度(du)(du)(du)(du)(du)值(zhi)。大鼠低密(mi)(mi)(mi)(mi)度(du)(du)(du)(du)(du)脂(zhi)(zhi)(zhi)蛋(dan)白(bai)(bai)免(mian)(mian)疫復(fu)(fu)合(he)物(wu)(wu)(wu)(wu)(wu)(LDL-IC) ELISA 試(shi)(shi)(shi)劑(ji)(ji)盒(he) 試(shi)(shi)(shi)劑(ji)(ji)盒(he)性(xing)(xing)(xing)能1.準(zhun)(zhun)(zhun)(zhun)確性(xing)(xing)(xing)：標(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)(pin)(pin)線(xian)性(xing)(xing)(xing)回(hui)(hui)歸(gui)與預期(qi)濃(nong)(nong)度(du)(du)(du)(du)(du)相關系數(shu)(shu)R值(zhi)，大于(yu)等于(yu)0.9900。2.靈敏度(du)(du)(du)(du)(du)：＊低檢(jian)測(ce)濃(nong)(nong)度(du)(du)(du)(du)(du)小(xiao)于(yu)1.0 U/ml。3.特異(yi)性(xing)(xing)(xing)：不(bu)(bu)與其它可(ke)(ke)溶性(xing)(xing)(xing)結構類似物(wu)(wu)(wu)(wu)(wu)交叉反應(ying)。4.重(zhong)復(fu)(fu)性(xing)(xing)(xing)：板(ban)(ban)(ban)內、板(ban)(ban)(ban)間(jian)變異(yi)系數(shu)(shu)均(jun)(jun)小(xiao)于(yu)15%。5.貯藏(zang)：2-8℃，避光防潮保存。6.有效期(qi)：6個月免(mian)(mian)責聲明(ming)(ming)(ming)1.試(shi)(shi)(shi)劑(ji)(ji)盒(he)僅(jin)供(gong)研究使(shi)用(yong)(yong)(yong)，不(bu)(bu)得用(yong)(yong)(yong)于(yu)臨床實(shi)(shi)驗或人(ren)體(ti)(ti)(ti)實(shi)(shi)驗，否則所(suo)產生的(de)(de)(de)(de)(de)(de)(de)一(yi)切后(hou)果，由(you)實(shi)(shi)驗者(zhe)承擔，本(ben)(ben)(ben)公司概不(bu)(bu)負責。2.嚴格按(an)照(zhao)(zhao)說(shuo)(shuo)明(ming)(ming)(ming)書(shu)操(cao)(cao)(cao)作(zuo)(zuo)(zuo)，實(shi)(shi)驗者(zhe)違反說(shuo)(shuo)明(ming)(ming)(ming)書(shu)操(cao)(cao)(cao)作(zuo)(zuo)(zuo)，后(hou)果由(you)實(shi)(shi)驗者(zhe)承擔。 大鼠低密(mi)(mi)(mi)(mi)度(du)(du)(du)(du)(du)脂(zhi)(zhi)(zhi)蛋(dan)白(bai)(bai)免(mian)(mian)疫復(fu)(fu)合(he)物(wu)(wu)(wu)(wu)(wu)(LDL-IC) ELISA 試(shi)(shi)(shi)劑(ji)(ji)盒(he)
大(da)鼠(shu)(shu)低(di)(di)密(mi)(mi)度(du)(du)(du)(du)脂蛋(dan)(dan)(dan)(dan)白(bai)(bai)受(shou)體(ti)(ti)(ti)(ti)相(xiang)(xiang)關(guan)(guan)蛋(dan)(dan)(dan)(dan)白(bai)(bai)1(LRP-1) ELISA 試(shi)(shi)(shi)劑(ji)盒(he)(he)(he) 大(da)鼠(shu)(shu)低(di)(di)密(mi)(mi)度(du)(du)(du)(du)脂蛋(dan)(dan)(dan)(dan)白(bai)(bai)受(shou)體(ti)(ti)(ti)(ti)相(xiang)(xiang)關(guan)(guan)蛋(dan)(dan)(dan)(dan)白(bai)(bai)1(LRP-1) ELISA 試(shi)(shi)(shi)劑(ji)盒(he)(he)(he) 檢(jian)測(ce)(ce)原(yuan)理試(shi)(shi)(shi)劑(ji)盒(he)(he)(he)采(cai)用(yong)(yong)(yong)(yong)雙(shuang)抗體(ti)(ti)(ti)(ti)一步夾心(xin)(xin)(xin)(xin)法酶(mei)(mei)聯免(mian)疫吸(xi)(xi)附試(shi)(shi)(shi)驗（ELISA）。往預先包(bao)被(bei)活性(xing)(xing)(xing)氧簇（ROS）抗體(ti)(ti)(ti)(ti)的(de)(de)(de)(de)(de)(de)包(bao)被(bei)微孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)中(zhong)，依(yi)(yi)次加(jia)(jia)(jia)(jia)(jia)入標(biao)(biao)(biao)(biao)本(ben)(ben)(ben)、標(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)(pin)、HRP標(biao)(biao)(biao)(biao)記(ji)的(de)(de)(de)(de)(de)(de)檢(jian)測(ce)(ce)抗體(ti)(ti)(ti)(ti)，經(jing)過(guo)溫(wen)(wen)(wen)(wen)(wen)育并徹底洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)。用(yong)(yong)(yong)(yong)底物(wu)(wu)(wu)(wu)TMB顯(xian)色，TMB在(zai)(zai)(zai)(zai)(zai)過(guo)氧化物(wu)(wu)(wu)(wu)酶(mei)(mei)的(de)(de)(de)(de)(de)(de)催(cui)化下(xia)轉(zhuan)(zhuan)(zhuan)化成藍色，并在(zai)(zai)(zai)(zai)(zai)酸(suan)的(de)(de)(de)(de)(de)(de)作(zuo)(zuo)(zuo)用(yong)(yong)(yong)(yong)下(xia)轉(zhuan)(zhuan)(zhuan)化成＊終(zhong)的(de)(de)(de)(de)(de)(de)黃色。顏色的(de)(de)(de)(de)(de)(de)深淺和(he)樣(yang)(yang)(yang)(yang)品(pin)(pin)(pin)(pin)(pin)(pin)中(zhong)的(de)(de)(de)(de)(de)(de)活性(xing)(xing)(xing)氧簇（ROS）呈正相(xiang)(xiang)關(guan)(guan)。用(yong)(yong)(yong)(yong)酶(mei)(mei)標(biao)(biao)(biao)(biao)儀(yi)在(zai)(zai)(zai)(zai)(zai)450nm 波長下(xia)測(ce)(ce)定(ding)吸(xi)(xi)光(guang)度(du)(du)(du)(du)（OD 值(zhi)），計算樣(yang)(yang)(yang)(yang)品(pin)(pin)(pin)(pin)(pin)(pin)活性(xing)(xing)(xing)。大(da)鼠(shu)(shu)低(di)(di)密(mi)(mi)度(du)(du)(du)(du)脂蛋(dan)(dan)(dan)(dan)白(bai)(bai)受(shou)體(ti)(ti)(ti)(ti)相(xiang)(xiang)關(guan)(guan)蛋(dan)(dan)(dan)(dan)白(bai)(bai)1(LRP-1) ELISA 試(shi)(shi)(shi)劑(ji)盒(he)(he)(he) 樣(yang)(yang)(yang)(yang)品(pin)(pin)(pin)(pin)(pin)(pin)收(shou)(shou)集、處理及保存(cun)方(fang)法1. 血(xue)(xue)清(qing)(qing)：使(shi)用(yong)(yong)(yong)(yong)不(bu)含熱(re)原(yuan)和(he)內(nei)毒素的(de)(de)(de)(de)(de)(de)試(shi)(shi)(shi)管(guan)(guan)，操作(zuo)(zuo)(zuo)過(guo)程(cheng)中(zhong)避免(mian)任(ren)何細(xi)胞刺激，收(shou)(shou)集血(xue)(xue)液(ye)(ye)(ye)(ye)后(hou)(hou)，3000轉(zhuan)(zhuan)(zhuan)離(li)(li)心(xin)(xin)(xin)(xin)10分(fen)(fen)(fen)鐘(zhong)將血(xue)(xue)清(qing)(qing)和(he)紅細(xi)胞迅(xun)速小(xiao)心(xin)(xin)(xin)(xin)地分(fen)(fen)(fen)離(li)(li)。2. 血(xue)(xue)漿(jiang)：EDTA、檸(ning)檬酸(suan)鹽(yan)或肝(gan)素抗凝。3000轉(zhuan)(zhuan)(zhuan)離(li)(li)心(xin)(xin)(xin)(xin)30分(fen)(fen)(fen)鐘(zhong)取(qu)上(shang)(shang)清(qing)(qing)。3. 細(xi)胞上(shang)(shang)清(qing)(qing)液(ye)(ye)(ye)(ye)：3000轉(zhuan)(zhuan)(zhuan)離(li)(li)心(xin)(xin)(xin)(xin)10分(fen)(fen)(fen)鐘(zhong)去除(chu)顆粒和(he)聚(ju)合物(wu)(wu)(wu)(wu)。4. 組織勻漿(jiang)：將組織加(jia)(jia)(jia)(jia)(jia)入適(shi)量生理鹽(yan)水搗碎(sui)。3000轉(zhuan)(zhuan)(zhuan)離(li)(li)心(xin)(xin)(xin)(xin)10分(fen)(fen)(fen)鐘(zhong)取(qu)上(shang)(shang)清(qing)(qing)。5. 保存(cun)：如(ru)果(guo)(guo)樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)收(shou)(shou)集后(hou)(hou)不(bu)及時(shi)檢(jian)測(ce)(ce)，請按(an)(an)一次用(yong)(yong)(yong)(yong)量分(fen)(fen)(fen)裝，凍(dong)(dong)存(cun)于-20℃，避免(mian)反(fan)(fan)復(fu)凍(dong)(dong)融，在(zai)(zai)(zai)(zai)(zai)室溫(wen)(wen)(wen)(wen)(wen)下(xia)解凍(dong)(dong)并確保樣(yang)(yang)(yang)(yang)品(pin)(pin)(pin)(pin)(pin)(pin)均(jun)勻地充(chong)分(fen)(fen)(fen)解凍(dong)(dong)。大(da)鼠(shu)(shu)低(di)(di)密(mi)(mi)度(du)(du)(du)(du)脂蛋(dan)(dan)(dan)(dan)白(bai)(bai)受(shou)體(ti)(ti)(ti)(ti)相(xiang)(xiang)關(guan)(guan)蛋(dan)(dan)(dan)(dan)白(bai)(bai)1(LRP-1) ELISA 試(shi)(shi)(shi)劑(ji)盒(he)(he)(he) 自備物(wu)(wu)(wu)(wu)品(pin)(pin)(pin)(pin)(pin)(pin)1.酶(mei)(mei)標(biao)(biao)(biao)(biao)儀(yi)（450nm）2.高(gao)精度(du)(du)(du)(du)加(jia)(jia)(jia)(jia)(jia)樣(yang)(yang)(yang)(yang)器(qi)及槍(qiang)頭：0.5-10uL、2-20uL、20-200uL、200-1000uL3.37℃恒溫(wen)(wen)(wen)(wen)(wen)箱操作(zuo)(zuo)(zuo)注(zhu)(zhu)意事(shi)項1.試(shi)(shi)(shi)劑(ji)盒(he)(he)(he)保存(cun)在(zai)(zai)(zai)(zai)(zai)2-8℃，使(shi)用(yong)(yong)(yong)(yong)前室溫(wen)(wen)(wen)(wen)(wen)平衡20分(fen)(fen)(fen)鐘(zhong)。從冰箱取(qu)出的(de)(de)(de)(de)(de)(de)濃(nong)縮洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)液(ye)(ye)(ye)(ye)會有結(jie)晶，這屬于正常現象，水浴加(jia)(jia)(jia)(jia)(jia)熱(re)使(shi)結(jie)晶完全溶解后(hou)(hou)再使(shi)用(yong)(yong)(yong)(yong)。2.實(shi)(shi)(shi)(shi)驗中(zhong)不(bu)用(yong)(yong)(yong)(yong)的(de)(de)(de)(de)(de)(de)板(ban)(ban)條(tiao)應立即(ji)放回自封(feng)(feng)(feng)袋(dai)(dai)中(zhong)，密(mi)(mi)封(feng)(feng)(feng)（低(di)(di)溫(wen)(wen)(wen)(wen)(wen)干(gan)(gan)燥）保存(cun)。3.濃(nong)度(du)(du)(du)(du)為0的(de)(de)(de)(de)(de)(de)S0號標(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)(pin)即(ji)可視為陰性(xing)(xing)(xing)對照或者空白(bai)(bai)；按(an)(an)照說(shuo)(shuo)明(ming)(ming)書操作(zuo)(zuo)(zuo)時(shi)樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)已經(jing)稀(xi)釋(shi)5倍，＊終(zhong)結(jie)果(guo)(guo)乘以5才(cai)是樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)實(shi)(shi)(shi)(shi)際(ji)濃(nong)度(du)(du)(du)(du)。4.嚴(yan)格(ge)按(an)(an)照說(shuo)(shuo)明(ming)(ming)書中(zhong)標(biao)(biao)(biao)(biao)明(ming)(ming)的(de)(de)(de)(de)(de)(de)時(shi)間、加(jia)(jia)(jia)(jia)(jia)液(ye)(ye)(ye)(ye)量及順序進(jin)行(xing)(xing)溫(wen)(wen)(wen)(wen)(wen)育操作(zuo)(zuo)(zuo)。5.所(suo)有液(ye)(ye)(ye)(ye)體(ti)(ti)(ti)(ti)組分(fen)(fen)(fen)使(shi)用(yong)(yong)(yong)(yong)前充(chong)分(fen)(fen)(fen)搖勻。試(shi)(shi)(shi)劑(ji)盒(he)(he)(he)組成名稱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(zhu)(zhu)微孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)酶(mei)(mei)標(biao)(biao)(biao)(biao)板(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)標(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)(pin)0.3mL*6管(guan)(guan)0.3mL*6管(guan)(guan)無(wu)樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)稀(xi)釋(shi)液(ye)(ye)(ye)(ye)6mL3mL無(wu)檢(jian)測(ce)(ce)抗體(ti)(ti)(ti)(ti)-HRP10mL5mL無(wu)20×洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)緩(huan)(huan)沖(chong)液(ye)(ye)(ye)(ye)25mL15mL按(an)(an)說(shuo)(shuo)明(ming)(ming)書進(jin)行(xing)(xing)稀(xi)釋(shi)底物(wu)(wu)(wu)(wu)A6mL3mL無(wu)底物(wu)(wu)(wu)(wu)B6mL3mL無(wu)終(zhong)止液(ye)(ye)(ye)(ye)6mL3mL無(wu)封(feng)(feng)(feng)板(ban)(ban)膜(mo)2張2張無(wu)說(shuo)(shuo)明(ming)(ming)書1份1份無(wu)自封(feng)(feng)(feng)袋(dai)(dai)1個(ge)(ge)1個(ge)(ge)無(wu)注(zhu)(zhu)：標(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)(pin)（S0-S5）濃(nong)度(du)(du)(du)(du)依(yi)(yi)次為：0、30、60、120、240、480 U/ml試(shi)(shi)(shi)劑(ji)的(de)(de)(de)(de)(de)(de)準(zhun)(zhun)(zhun)備 20×洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)緩(huan)(huan)沖(chong)液(ye)(ye)(ye)(ye)的(de)(de)(de)(de)(de)(de)稀(xi)釋(shi)：蒸(zheng)餾(liu)水按(an)(an)1：20稀(xi)釋(shi)，即(ji)1份的(de)(de)(de)(de)(de)(de)20×洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)緩(huan)(huan)沖(chong)液(ye)(ye)(ye)(ye)加(jia)(jia)(jia)(jia)(jia)19份的(de)(de)(de)(de)(de)(de)蒸(zheng)餾(liu)水。洗(xi)(xi)(xi)(xi)(xi)板(ban)(ban)方(fang)法1.手(shou)工洗(xi)(xi)(xi)(xi)(xi)板(ban)(ban)：甩(shuai)盡孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)內(nei)液(ye)(ye)(ye)(ye)體(ti)(ti)(ti)(ti)，每孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)(jia)(jia)(jia)滿洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)液(ye)(ye)(ye)(ye)，靜置(zhi)1min后(hou)(hou)甩(shuai)盡孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)內(nei)液(ye)(ye)(ye)(ye)體(ti)(ti)(ti)(ti)，在(zai)(zai)(zai)(zai)(zai)吸(xi)(xi)水紙(zhi)上(shang)(shang)拍干(gan)(gan)，如(ru)此洗(xi)(xi)(xi)(xi)(xi)板(ban)(ban)5次。2.自動洗(xi)(xi)(xi)(xi)(xi)板(ban)(ban)機(ji)：每孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)注(zhu)(zhu)入洗(xi)(xi)(xi)(xi)(xi)液(ye)(ye)(ye)(ye)350μL，浸泡1min，洗(xi)(xi)(xi)(xi)(xi)板(ban)(ban)5次。操作(zuo)(zuo)(zuo)步驟1.從室溫(wen)(wen)(wen)(wen)(wen)平衡20min后(hou)(hou)的(de)(de)(de)(de)(de)(de)鋁箔(bo)袋(dai)(dai)中(zhong)取(qu)出所(suo)需板(ban)(ban)條(tiao)，剩(sheng)余板(ban)(ban)條(tiao)用(yong)(yong)(yong)(yong)自封(feng)(feng)(feng)袋(dai)(dai)密(mi)(mi)封(feng)(feng)(feng)放回4℃。2.設置(zhi)標(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)(pin)孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)和(he)樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)，標(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)(pin)孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)各(ge)加(jia)(jia)(jia)(jia)(jia)不(bu)同(tong)濃(nong)度(du)(du)(du)(du)的(de)(de)(de)(de)(de)(de)標(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)(pin)50μL；3.樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)先加(jia)(jia)(jia)(jia)(jia)待(dai)測(ce)(ce)樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)10μL，再加(jia)(jia)(jia)(jia)(jia)樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)稀(xi)釋(shi)液(ye)(ye)(ye)(ye)40μL；空白(bai)(bai)孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)不(bu)加(jia)(jia)(jia)(jia)(jia)。4.除(chu)空白(bai)(bai)孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)外，標(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)(pin)孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)和(he)樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)中(zhong)每孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)(jia)(jia)(jia)入辣根(gen)過(guo)氧化物(wu)(wu)(wu)(wu)酶(mei)(mei)（HRP）標(biao)(biao)(biao)(biao)記(ji)的(de)(de)(de)(de)(de)(de)檢(jian)測(ce)(ce)抗體(ti)(ti)(ti)(ti)100μL，用(yong)(yong)(yong)(yong)封(feng)(feng)(feng)板(ban)(ban)膜(mo)封(feng)(feng)(feng)住反(fan)(fan)應孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)，37℃水浴鍋或恒溫(wen)(wen)(wen)(wen)(wen)箱溫(wen)(wen)(wen)(wen)(wen)育60min。5.棄去液(ye)(ye)(ye)(ye)體(ti)(ti)(ti)(ti)，吸(xi)(xi)水紙(zhi)上(shang)(shang)拍干(gan)(gan)，每孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)(jia)(jia)(jia)滿洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)液(ye)(ye)(ye)(ye)，靜置(zhi)1min，甩(shuai)去洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)液(ye)(ye)(ye)(ye)，吸(xi)(xi)水紙(zhi)上(shang)(shang)拍干(gan)(gan)，如(ru)此重復(fu)洗(xi)(xi)(xi)(xi)(xi)板(ban)(ban)5次（也可用(yong)(yong)(yong)(yong)洗(xi)(xi)(xi)(xi)(xi)板(ban)(ban)機(ji)洗(xi)(xi)(xi)(xi)(xi)板(ban)(ban)）。6.每孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)(jia)(jia)(jia)入底物(wu)(wu)(wu)(wu)A、B各(ge)50μL，37℃避光(guang)孵育15min。7.每孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)(jia)(jia)(jia)入終(zhong)止液(ye)(ye)(ye)(ye)50μL，15min內(nei)，在(zai)(zai)(zai)(zai)(zai)450nm波長處測(ce)(ce)定(ding)各(ge)孔(kong)(kong)(kong)(kong)(kong)(kong)(kong)的(de)(de)(de)(de)(de)(de)OD值(zhi)。結(jie)果(guo)(guo)判斷 繪制標(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)曲線：在(zai)(zai)(zai)(zai)(zai)Excel工作(zuo)(zuo)(zuo)表中(zhong)，以標(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)(pin)濃(nong)度(du)(du)(du)(du)作(zuo)(zuo)(zuo)橫坐(zuo)標(biao)(biao)(biao)(biao)，對應OD值(zhi)作(zuo)(zuo)(zuo)縱坐(zuo)標(biao)(biao)(biao)(biao)，繪制出標(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)(pin)線性(xing)(xing)(xing)回歸(gui)(gui)曲線，按(an)(an)曲線方(fang)程(cheng)計算各(ge)樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)濃(nong)度(du)(du)(du)(du)值(zhi)。大(da)鼠(shu)(shu)低(di)(di)密(mi)(mi)度(du)(du)(du)(du)脂蛋(dan)(dan)(dan)(dan)白(bai)(bai)受(shou)體(ti)(ti)(ti)(ti)相(xiang)(xiang)關(guan)(guan)蛋(dan)(dan)(dan)(dan)白(bai)(bai)1(LRP-1) ELISA 試(shi)(shi)(shi)劑(ji)盒(he)(he)(he) 試(shi)(shi)(shi)劑(ji)盒(he)(he)(he)性(xing)(xing)(xing)能(neng)1.準(zhun)(zhun)(zhun)確性(xing)(xing)(xing)：標(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)(pin)(pin)(pin)(pin)(pin)線性(xing)(xing)(xing)回歸(gui)(gui)與(yu)(yu)預期濃(nong)度(du)(du)(du)(du)相(xiang)(xiang)關(guan)(guan)系數R值(zhi)，大(da)于等于0.9900。2.靈敏(min)度(du)(du)(du)(du)：＊低(di)(di)檢(jian)測(ce)(ce)濃(nong)度(du)(du)(du)(du)小(xiao)于1.0 U/ml。3.特異(yi)性(xing)(xing)(xing)：不(bu)與(yu)(yu)其它可溶性(xing)(xing)(xing)結(jie)構類似物(wu)(wu)(wu)(wu)交(jiao)叉(cha)反(fan)(fan)應。4.重復(fu)性(xing)(xing)(xing)：板(ban)(ban)內(nei)、板(ban)(ban)間變(bian)異(yi)系數均(jun)小(xiao)于15%。5.貯藏：2-8℃，避光(guang)防(fang)潮保存(cun)。6.有效期：6個(ge)(ge)月(yue)免(mian)責聲(sheng)明(ming)(ming)1.試(shi)(shi)(shi)劑(ji)盒(he)(he)(he)僅供(gong)研究(jiu)使(shi)用(yong)(yong)(yong)(yong)，不(bu)得用(yong)(yong)(yong)(yong)于臨床實(shi)(shi)(shi)(shi)驗或人體(ti)(ti)(ti)(ti)實(shi)(shi)(shi)(shi)驗，否(fou)則(ze)所(suo)產生的(de)(de)(de)(de)(de)(de)一切后(hou)(hou)果(guo)(guo)，由實(shi)(shi)(shi)(shi)驗者承擔，本(ben)(ben)(ben)公司概不(bu)負責。2.嚴(yan)格(ge)按(an)(an)照說(shuo)(shuo)明(ming)(ming)書操作(zuo)(zuo)(zuo)，實(shi)(shi)(shi)(shi)驗者違反(fan)(fan)說(shuo)(shuo)明(ming)(ming)書操作(zuo)(zuo)(zuo)，后(hou)(hou)果(guo)(guo)由實(shi)(shi)(shi)(shi)驗者承擔。 大(da)鼠(shu)(shu)低(di)(di)密(mi)(mi)度(du)(du)(du)(du)脂蛋(dan)(dan)(dan)(dan)白(bai)(bai)受(shou)體(ti)(ti)(ti)(ti)相(xiang)(xiang)關(guan)(guan)蛋(dan)(dan)(dan)(dan)白(bai)(bai)1(LRP-1) ELISA 試(shi)(shi)(shi)劑(ji)盒(he)(he)(he)
大(da)鼠低密(mi)(mi)度(du)(du)(du)脂(zhi)(zhi)(zhi)蛋(dan)白(bai)(bai)(LDL) ELISA 試(shi)(shi)(shi)劑(ji)盒(he)(he)(he) 大(da)鼠低密(mi)(mi)度(du)(du)(du)脂(zhi)(zhi)(zhi)蛋(dan)白(bai)(bai)(LDL) ELISA 試(shi)(shi)(shi)劑(ji)盒(he)(he)(he) 檢(jian)(jian)測(ce)(ce)原(yuan)理試(shi)(shi)(shi)劑(ji)盒(he)(he)(he)采(cai)用(yong)雙抗(kang)體(ti)(ti)(ti)(ti)(ti)一(yi)步夾心法(fa)(fa)酶(mei)聯免(mian)疫吸(xi)(xi)附試(shi)(shi)(shi)驗(yan)（ELISA）。往預先(xian)包被活性(xing)(xing)(xing)氧(yang)(yang)簇(cu)（ROS）抗(kang)體(ti)(ti)(ti)(ti)(ti)的(de)(de)包被微孔(kong)中(zhong)，依(yi)次加(jia)(jia)(jia)入(ru)標(biao)(biao)(biao)(biao)(biao)(biao)本(ben)(ben)(ben)(ben)(ben)、標(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)、HRP標(biao)(biao)(biao)(biao)(biao)(biao)記的(de)(de)檢(jian)(jian)測(ce)(ce)抗(kang)體(ti)(ti)(ti)(ti)(ti)，經(jing)過(guo)(guo)(guo)溫(wen)(wen)(wen)(wen)育(yu)(yu)并(bing)(bing)(bing)徹底洗(xi)(xi)(xi)滌(di)(di)(di)(di)(di)。用(yong)底物(wu)(wu)(wu)TMB顯(xian)色，TMB在(zai)(zai)(zai)過(guo)(guo)(guo)氧(yang)(yang)化(hua)(hua)物(wu)(wu)(wu)酶(mei)的(de)(de)催(cui)化(hua)(hua)下(xia)轉(zhuan)(zhuan)(zhuan)化(hua)(hua)成藍(lan)色，并(bing)(bing)(bing)在(zai)(zai)(zai)酸的(de)(de)作(zuo)用(yong)下(xia)轉(zhuan)(zhuan)(zhuan)化(hua)(hua)成＊終(zhong)(zhong)的(de)(de)黃色。顏色的(de)(de)深(shen)淺和(he)樣(yang)(yang)(yang)(yang)品(pin)中(zhong)的(de)(de)活性(xing)(xing)(xing)氧(yang)(yang)簇(cu)（ROS）呈正相(xiang)(xiang)關。用(yong)酶(mei)標(biao)(biao)(biao)(biao)(biao)(biao)儀(yi)在(zai)(zai)(zai)450nm 波(bo)(bo)長下(xia)測(ce)(ce)定吸(xi)(xi)光度(du)(du)(du)（OD 值(zhi)），計算樣(yang)(yang)(yang)(yang)品(pin)活性(xing)(xing)(xing)。大(da)鼠低密(mi)(mi)度(du)(du)(du)脂(zhi)(zhi)(zhi)蛋(dan)白(bai)(bai)(LDL) ELISA 試(shi)(shi)(shi)劑(ji)盒(he)(he)(he) 樣(yang)(yang)(yang)(yang)品(pin)收(shou)集(ji)、處(chu)理及(ji)保存方法(fa)(fa)1. 血清(qing)(qing)：使(shi)(shi)(shi)(shi)用(yong)不(bu)含熱(re)原(yuan)和(he)內(nei)毒素的(de)(de)試(shi)(shi)(shi)管(guan)，操(cao)作(zuo)過(guo)(guo)(guo)程中(zhong)避(bi)(bi)免(mian)任何細(xi)胞(bao)刺激，收(shou)集(ji)血液(ye)后(hou)，3000轉(zhuan)(zhuan)(zhuan)離心10分(fen)(fen)鐘將(jiang)血清(qing)(qing)和(he)紅(hong)細(xi)胞(bao)迅速小心地分(fen)(fen)離。2. 血漿(jiang)(jiang)：EDTA、檸檬(meng)酸鹽或(huo)(huo)肝素抗(kang)凝。3000轉(zhuan)(zhuan)(zhuan)離心30分(fen)(fen)鐘取上(shang)(shang)(shang)清(qing)(qing)。3. 細(xi)胞(bao)上(shang)(shang)(shang)清(qing)(qing)液(ye)：3000轉(zhuan)(zhuan)(zhuan)離心10分(fen)(fen)鐘去(qu)除顆粒和(he)聚合(he)物(wu)(wu)(wu)。4. 組(zu)織勻(yun)漿(jiang)(jiang)：將(jiang)組(zu)織加(jia)(jia)(jia)入(ru)適量(liang)生理鹽水(shui)搗碎。3000轉(zhuan)(zhuan)(zhuan)離心10分(fen)(fen)鐘取上(shang)(shang)(shang)清(qing)(qing)。5. 保存：如(ru)果(guo)(guo)樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)收(shou)集(ji)后(hou)不(bu)及(ji)時(shi)檢(jian)(jian)測(ce)(ce)，請按(an)(an)一(yi)次用(yong)量(liang)分(fen)(fen)裝，凍存于(yu)-20℃，避(bi)(bi)免(mian)反復(fu)(fu)凍融，在(zai)(zai)(zai)室溫(wen)(wen)(wen)(wen)下(xia)解(jie)凍并(bing)(bing)(bing)確保樣(yang)(yang)(yang)(yang)品(pin)均勻(yun)地充(chong)分(fen)(fen)解(jie)凍。大(da)鼠低密(mi)(mi)度(du)(du)(du)脂(zhi)(zhi)(zhi)蛋(dan)白(bai)(bai)(LDL) ELISA 試(shi)(shi)(shi)劑(ji)盒(he)(he)(he) 自(zi)備(bei)物(wu)(wu)(wu)品(pin)1.酶(mei)標(biao)(biao)(biao)(biao)(biao)(biao)儀(yi)（450nm）2.高精(jing)度(du)(du)(du)加(jia)(jia)(jia)樣(yang)(yang)(yang)(yang)器及(ji)槍頭(tou)：0.5-10uL、2-20uL、20-200uL、200-1000uL3.37℃恒(heng)溫(wen)(wen)(wen)(wen)箱(xiang)(xiang)操(cao)作(zuo)注意(yi)事項1.試(shi)(shi)(shi)劑(ji)盒(he)(he)(he)保存在(zai)(zai)(zai)2-8℃，使(shi)(shi)(shi)(shi)用(yong)前室溫(wen)(wen)(wen)(wen)平(ping)衡20分(fen)(fen)鐘。從(cong)冰(bing)箱(xiang)(xiang)取出的(de)(de)濃(nong)(nong)(nong)縮洗(xi)(xi)(xi)滌(di)(di)(di)(di)(di)液(ye)會有(you)結晶(jing)，這屬(shu)于(yu)正常現象，水(shui)浴加(jia)(jia)(jia)熱(re)使(shi)(shi)(shi)(shi)結晶(jing)完全溶(rong)解(jie)后(hou)再使(shi)(shi)(shi)(shi)用(yong)。2.實(shi)驗(yan)中(zhong)不(bu)用(yong)的(de)(de)板(ban)(ban)(ban)條應立即放回自(zi)封(feng)(feng)(feng)袋(dai)中(zhong)，密(mi)(mi)封(feng)(feng)(feng)（低溫(wen)(wen)(wen)(wen)干燥）保存。3.濃(nong)(nong)(nong)度(du)(du)(du)為(wei)0的(de)(de)S0號(hao)標(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)即可(ke)視為(wei)陰性(xing)(xing)(xing)對照(zhao)(zhao)或(huo)(huo)者(zhe)空(kong)白(bai)(bai)；按(an)(an)照(zhao)(zhao)說明書(shu)操(cao)作(zuo)時(shi)樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)已經(jing)稀(xi)(xi)釋(shi)(shi)5倍，＊終(zhong)(zhong)結果(guo)(guo)乘以5才是樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)實(shi)際濃(nong)(nong)(nong)度(du)(du)(du)。4.嚴(yan)格按(an)(an)照(zhao)(zhao)說明書(shu)中(zhong)標(biao)(biao)(biao)(biao)(biao)(biao)明的(de)(de)時(shi)間、加(jia)(jia)(jia)液(ye)量(liang)及(ji)順(shun)序進(jin)行溫(wen)(wen)(wen)(wen)育(yu)(yu)操(cao)作(zuo)。5.所有(you)液(ye)體(ti)(ti)(ti)(ti)(ti)組(zu)分(fen)(fen)使(shi)(shi)(shi)(shi)用(yong)前充(chong)分(fen)(fen)搖勻(yun)。試(shi)(shi)(shi)劑(ji)盒(he)(he)(he)組(zu)成名(ming)稱96孔(kong)配(pei)置(zhi)48孔(kong)配(pei)置(zhi)備(bei)注微孔(kong)酶(mei)標(biao)(biao)(biao)(biao)(biao)(biao)板(ban)(ban)(ban)12孔(kong)×8條12孔(kong)×4條無(wu)(wu)(wu)標(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)0.3mL*6管(guan)0.3mL*6管(guan)無(wu)(wu)(wu)樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)稀(xi)(xi)釋(shi)(shi)液(ye)6mL3mL無(wu)(wu)(wu)檢(jian)(jian)測(ce)(ce)抗(kang)體(ti)(ti)(ti)(ti)(ti)-HRP10mL5mL無(wu)(wu)(wu)20×洗(xi)(xi)(xi)滌(di)(di)(di)(di)(di)緩沖(chong)液(ye)25mL15mL按(an)(an)說明書(shu)進(jin)行稀(xi)(xi)釋(shi)(shi)底物(wu)(wu)(wu)A6mL3mL無(wu)(wu)(wu)底物(wu)(wu)(wu)B6mL3mL無(wu)(wu)(wu)終(zhong)(zhong)止液(ye)6mL3mL無(wu)(wu)(wu)封(feng)(feng)(feng)板(ban)(ban)(ban)膜(mo)(mo)2張(zhang)2張(zhang)無(wu)(wu)(wu)說明書(shu)1份(fen)1份(fen)無(wu)(wu)(wu)自(zi)封(feng)(feng)(feng)袋(dai)1個(ge)1個(ge)無(wu)(wu)(wu)注：標(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)（S0-S5）濃(nong)(nong)(nong)度(du)(du)(du)依(yi)次為(wei)：0、30、60、120、240、480 U/ml試(shi)(shi)(shi)劑(ji)的(de)(de)準(zhun)(zhun)(zhun)(zhun)備(bei) 20×洗(xi)(xi)(xi)滌(di)(di)(di)(di)(di)緩沖(chong)液(ye)的(de)(de)稀(xi)(xi)釋(shi)(shi)：蒸(zheng)餾水(shui)按(an)(an)1：20稀(xi)(xi)釋(shi)(shi)，即1份(fen)的(de)(de)20×洗(xi)(xi)(xi)滌(di)(di)(di)(di)(di)緩沖(chong)液(ye)加(jia)(jia)(jia)19份(fen)的(de)(de)蒸(zheng)餾水(shui)。洗(xi)(xi)(xi)板(ban)(ban)(ban)方法(fa)(fa)1.手工洗(xi)(xi)(xi)板(ban)(ban)(ban)：甩盡孔(kong)內(nei)液(ye)體(ti)(ti)(ti)(ti)(ti)，每孔(kong)加(jia)(jia)(jia)滿(man)洗(xi)(xi)(xi)滌(di)(di)(di)(di)(di)液(ye)，靜(jing)置(zhi)1min后(hou)甩盡孔(kong)內(nei)液(ye)體(ti)(ti)(ti)(ti)(ti)，在(zai)(zai)(zai)吸(xi)(xi)水(shui)紙上(shang)(shang)(shang)拍干，如(ru)此洗(xi)(xi)(xi)板(ban)(ban)(ban)5次。2.自(zi)動洗(xi)(xi)(xi)板(ban)(ban)(ban)機：每孔(kong)注入(ru)洗(xi)(xi)(xi)液(ye)350μL，浸泡1min，洗(xi)(xi)(xi)板(ban)(ban)(ban)5次。操(cao)作(zuo)步驟(zou)1.從(cong)室溫(wen)(wen)(wen)(wen)平(ping)衡20min后(hou)的(de)(de)鋁箔袋(dai)中(zhong)取出所需板(ban)(ban)(ban)條，剩余板(ban)(ban)(ban)條用(yong)自(zi)封(feng)(feng)(feng)袋(dai)密(mi)(mi)封(feng)(feng)(feng)放回4℃。2.設置(zhi)標(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)孔(kong)和(he)樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)孔(kong)，標(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)孔(kong)各(ge)(ge)加(jia)(jia)(jia)不(bu)同濃(nong)(nong)(nong)度(du)(du)(du)的(de)(de)標(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)50μL；3.樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)孔(kong)先(xian)加(jia)(jia)(jia)待測(ce)(ce)樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)10μL，再加(jia)(jia)(jia)樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)稀(xi)(xi)釋(shi)(shi)液(ye)40μL；空(kong)白(bai)(bai)孔(kong)不(bu)加(jia)(jia)(jia)。4.除空(kong)白(bai)(bai)孔(kong)外(wai)，標(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)孔(kong)和(he)樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)孔(kong)中(zhong)每孔(kong)加(jia)(jia)(jia)入(ru)辣根過(guo)(guo)(guo)氧(yang)(yang)化(hua)(hua)物(wu)(wu)(wu)酶(mei)（HRP）標(biao)(biao)(biao)(biao)(biao)(biao)記的(de)(de)檢(jian)(jian)測(ce)(ce)抗(kang)體(ti)(ti)(ti)(ti)(ti)100μL，用(yong)封(feng)(feng)(feng)板(ban)(ban)(ban)膜(mo)(mo)封(feng)(feng)(feng)住反應孔(kong)，37℃水(shui)浴鍋或(huo)(huo)恒(heng)溫(wen)(wen)(wen)(wen)箱(xiang)(xiang)溫(wen)(wen)(wen)(wen)育(yu)(yu)60min。5.棄去(qu)液(ye)體(ti)(ti)(ti)(ti)(ti)，吸(xi)(xi)水(shui)紙上(shang)(shang)(shang)拍干，每孔(kong)加(jia)(jia)(jia)滿(man)洗(xi)(xi)(xi)滌(di)(di)(di)(di)(di)液(ye)，靜(jing)置(zhi)1min，甩去(qu)洗(xi)(xi)(xi)滌(di)(di)(di)(di)(di)液(ye)，吸(xi)(xi)水(shui)紙上(shang)(shang)(shang)拍干，如(ru)此重復(fu)(fu)洗(xi)(xi)(xi)板(ban)(ban)(ban)5次（也可(ke)用(yong)洗(xi)(xi)(xi)板(ban)(ban)(ban)機洗(xi)(xi)(xi)板(ban)(ban)(ban)）。6.每孔(kong)加(jia)(jia)(jia)入(ru)底物(wu)(wu)(wu)A、B各(ge)(ge)50μL，37℃避(bi)(bi)光孵育(yu)(yu)15min。7.每孔(kong)加(jia)(jia)(jia)入(ru)終(zhong)(zhong)止液(ye)50μL，15min內(nei)，在(zai)(zai)(zai)450nm波(bo)(bo)長處(chu)測(ce)(ce)定各(ge)(ge)孔(kong)的(de)(de)OD值(zhi)。結果(guo)(guo)判斷 繪制標(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)曲(qu)線：在(zai)(zai)(zai)Excel工作(zuo)表(biao)中(zhong)，以標(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)濃(nong)(nong)(nong)度(du)(du)(du)作(zuo)橫坐(zuo)標(biao)(biao)(biao)(biao)(biao)(biao)，對應OD值(zhi)作(zuo)縱坐(zuo)標(biao)(biao)(biao)(biao)(biao)(biao)，繪制出標(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品(pin)線性(xing)(xing)(xing)回歸(gui)(gui)曲(qu)線，按(an)(an)曲(qu)線方程計算各(ge)(ge)樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)濃(nong)(nong)(nong)度(du)(du)(du)值(zhi)。大(da)鼠低密(mi)(mi)度(du)(du)(du)脂(zhi)(zhi)(zhi)蛋(dan)白(bai)(bai)(LDL) ELISA 試(shi)(shi)(shi)劑(ji)盒(he)(he)(he) 試(shi)(shi)(shi)劑(ji)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)準(zhun)(zhun)(zhun)(zhun)品(pin)線性(xing)(xing)(xing)回歸(gui)(gui)與(yu)預期(qi)濃(nong)(nong)(nong)度(du)(du)(du)相(xiang)(xiang)關系數R值(zhi)，大(da)于(yu)等于(yu)0.9900。2.靈(ling)敏度(du)(du)(du)：＊低檢(jian)(jian)測(ce)(ce)濃(nong)(nong)(nong)度(du)(du)(du)小于(yu)1.0 U/ml。3.特異性(xing)(xing)(xing)：不(bu)與(yu)其(qi)它可(ke)溶(rong)性(xing)(xing)(xing)結構類似物(wu)(wu)(wu)交叉反應。4.重復(fu)(fu)性(xing)(xing)(xing)：板(ban)(ban)(ban)內(nei)、板(ban)(ban)(ban)間變異系數均小于(yu)15%。5.貯藏：2-8℃，避(bi)(bi)光防潮保存。6.有(you)效(xiao)期(qi)：6個(ge)月免(mian)責聲明1.試(shi)(shi)(shi)劑(ji)盒(he)(he)(he)僅供研究(jiu)使(shi)(shi)(shi)(shi)用(yong)，不(bu)得用(yong)于(yu)臨(lin)床實(shi)驗(yan)或(huo)(huo)人體(ti)(ti)(ti)(ti)(ti)實(shi)驗(yan)，否則所產生的(de)(de)一(yi)切后(hou)果(guo)(guo)，由實(shi)驗(yan)者(zhe)承(cheng)擔(dan)，本(ben)(ben)(ben)(ben)(ben)公司概(gai)不(bu)負責。2.嚴(yan)格按(an)(an)照(zhao)(zhao)說明書(shu)操(cao)作(zuo)，實(shi)驗(yan)者(zhe)違反說明書(shu)操(cao)作(zuo)，后(hou)果(guo)(guo)由實(shi)驗(yan)者(zhe)承(cheng)擔(dan)。 大(da)鼠低密(mi)(mi)度(du)(du)(du)脂(zhi)(zhi)(zhi)蛋(dan)白(bai)(bai)(LDL) ELISA 試(shi)(shi)(shi)劑(ji)盒(he)(he)(he)
大(da)鼠纖維(wei)連(lian)(lian)接(jie)(jie)蛋(dan)白(bai)(FN) ELISA 試(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)(he) 大(da)鼠纖維(wei)連(lian)(lian)接(jie)(jie)蛋(dan)白(bai)(FN) ELISA 試(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)(he) 檢測(ce)(ce)原理試(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)(he)采(cai)用(yong)雙抗(kang)(kang)體(ti)(ti)(ti)一(yi)步夾心法(fa)酶(mei)聯免(mian)疫(yi)吸(xi)附試(shi)(shi)(shi)驗(yan)(yan)（ELISA）。往預先(xian)包被活性(xing)(xing)氧(yang)簇（ROS）抗(kang)(kang)體(ti)(ti)(ti)的(de)(de)(de)(de)包被微孔(kong)(kong)(kong)(kong)中(zhong)(zhong)，依(yi)次加(jia)(jia)入標(biao)(biao)(biao)(biao)(biao)(biao)(biao)本(ben)、標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)、HRP標(biao)(biao)(biao)(biao)(biao)(biao)(biao)記的(de)(de)(de)(de)檢測(ce)(ce)抗(kang)(kang)體(ti)(ti)(ti)，經過溫(wen)(wen)育并(bing)徹(che)底(di)洗(xi)(xi)(xi)滌(di)(di)。用(yong)底(di)物(wu)TMB顯色(se)(se)，TMB在(zai)(zai)過氧(yang)化(hua)物(wu)酶(mei)的(de)(de)(de)(de)催(cui)化(hua)下轉(zhuan)化(hua)成藍色(se)(se)，并(bing)在(zai)(zai)酸的(de)(de)(de)(de)作(zuo)(zuo)用(yong)下轉(zhuan)化(hua)成＊終(zhong)的(de)(de)(de)(de)黃色(se)(se)。顏色(se)(se)的(de)(de)(de)(de)深淺和樣(yang)(yang)品(pin)中(zhong)(zhong)的(de)(de)(de)(de)活性(xing)(xing)氧(yang)簇（ROS）呈正相關。用(yong)酶(mei)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)儀在(zai)(zai)450nm 波長下測(ce)(ce)定(ding)吸(xi)光(guang)度(du)（OD 值(zhi)），計算樣(yang)(yang)品(pin)活性(xing)(xing)。大(da)鼠纖維(wei)連(lian)(lian)接(jie)(jie)蛋(dan)白(bai)(FN) ELISA 試(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)(he) 樣(yang)(yang)品(pin)收集、處理及(ji)保(bao)存方法(fa)1. 血清(qing)：使(shi)(shi)用(yong)不(bu)(bu)含(han)熱原和內(nei)(nei)毒素(su)的(de)(de)(de)(de)試(shi)(shi)(shi)管(guan)，操(cao)(cao)作(zuo)(zuo)過程(cheng)中(zhong)(zhong)避免(mian)任何細胞刺(ci)激(ji)，收集血液(ye)(ye)(ye)(ye)(ye)(ye)后(hou)，3000轉(zhuan)離(li)(li)心10分(fen)(fen)(fen)鐘(zhong)(zhong)將血清(qing)和紅細胞迅速小(xiao)(xiao)心地分(fen)(fen)(fen)離(li)(li)。2. 血漿：EDTA、檸檬酸鹽或(huo)肝(gan)素(su)抗(kang)(kang)凝。3000轉(zhuan)離(li)(li)心30分(fen)(fen)(fen)鐘(zhong)(zhong)取(qu)上(shang)清(qing)。3. 細胞上(shang)清(qing)液(ye)(ye)(ye)(ye)(ye)(ye)：3000轉(zhuan)離(li)(li)心10分(fen)(fen)(fen)鐘(zhong)(zhong)去(qu)除(chu)(chu)顆粒和聚合物(wu)。4. 組(zu)織(zhi)勻(yun)漿：將組(zu)織(zhi)加(jia)(jia)入適(shi)量(liang)生理鹽水(shui)(shui)搗碎。3000轉(zhuan)離(li)(li)心10分(fen)(fen)(fen)鐘(zhong)(zhong)取(qu)上(shang)清(qing)。5. 保(bao)存：如(ru)(ru)果(guo)樣(yang)(yang)本(ben)收集后(hou)不(bu)(bu)及(ji)時(shi)檢測(ce)(ce)，請按(an)一(yi)次用(yong)量(liang)分(fen)(fen)(fen)裝，凍存于(yu)-20℃，避免(mian)反(fan)復凍融，在(zai)(zai)室(shi)溫(wen)(wen)下解(jie)凍并(bing)確保(bao)樣(yang)(yang)品(pin)均勻(yun)地充分(fen)(fen)(fen)解(jie)凍。大(da)鼠纖維(wei)連(lian)(lian)接(jie)(jie)蛋(dan)白(bai)(FN) ELISA 試(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)(he) 自(zi)備(bei)(bei)物(wu)品(pin)1.酶(mei)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)儀（450nm）2.高精度(du)加(jia)(jia)樣(yang)(yang)器及(ji)槍頭：0.5-10uL、2-20uL、20-200uL、200-1000uL3.37℃恒(heng)溫(wen)(wen)箱(xiang)(xiang)(xiang)操(cao)(cao)作(zuo)(zuo)注(zhu)意事項1.試(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)(he)保(bao)存在(zai)(zai)2-8℃，使(shi)(shi)用(yong)前(qian)室(shi)溫(wen)(wen)平(ping)衡20分(fen)(fen)(fen)鐘(zhong)(zhong)。從(cong)冰箱(xiang)(xiang)(xiang)取(qu)出的(de)(de)(de)(de)濃(nong)縮洗(xi)(xi)(xi)滌(di)(di)液(ye)(ye)(ye)(ye)(ye)(ye)會有結(jie)(jie)(jie)晶，這屬(shu)于(yu)正常現(xian)象，水(shui)(shui)浴加(jia)(jia)熱使(shi)(shi)結(jie)(jie)(jie)晶完全溶解(jie)后(hou)再使(shi)(shi)用(yong)。2.實(shi)驗(yan)(yan)中(zhong)(zhong)不(bu)(bu)用(yong)的(de)(de)(de)(de)板(ban)(ban)(ban)條(tiao)應立即(ji)放回自(zi)封袋(dai)中(zhong)(zhong)，密(mi)封（低(di)溫(wen)(wen)干燥）保(bao)存。3.濃(nong)度(du)為(wei)0的(de)(de)(de)(de)S0號標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)即(ji)可(ke)(ke)視(shi)為(wei)陰性(xing)(xing)對照(zhao)或(huo)者(zhe)空(kong)(kong)白(bai)；按(an)照(zhao)說(shuo)(shuo)明(ming)(ming)(ming)(ming)書(shu)(shu)(shu)操(cao)(cao)作(zuo)(zuo)時(shi)樣(yang)(yang)本(ben)已經稀(xi)釋(shi)(shi)5倍，＊終(zhong)結(jie)(jie)(jie)果(guo)乘以5才是(shi)樣(yang)(yang)本(ben)實(shi)際濃(nong)度(du)。4.嚴格按(an)照(zhao)說(shuo)(shuo)明(ming)(ming)(ming)(ming)書(shu)(shu)(shu)中(zhong)(zhong)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)明(ming)(ming)(ming)(ming)的(de)(de)(de)(de)時(shi)間、加(jia)(jia)液(ye)(ye)(ye)(ye)(ye)(ye)量(liang)及(ji)順(shun)序進(jin)行(xing)溫(wen)(wen)育操(cao)(cao)作(zuo)(zuo)。5.所有液(ye)(ye)(ye)(ye)(ye)(ye)體(ti)(ti)(ti)組(zu)分(fen)(fen)(fen)使(shi)(shi)用(yong)前(qian)充分(fen)(fen)(fen)搖勻(yun)。試(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)(he)組(zu)成名稱(cheng)96孔(kong)(kong)(kong)(kong)配(pei)置48孔(kong)(kong)(kong)(kong)配(pei)置備(bei)(bei)注(zhu)微孔(kong)(kong)(kong)(kong)酶(mei)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)板(ban)(ban)(ban)12孔(kong)(kong)(kong)(kong)×8條(tiao)12孔(kong)(kong)(kong)(kong)×4條(tiao)無(wu)(wu)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)0.3mL*6管(guan)0.3mL*6管(guan)無(wu)(wu)樣(yang)(yang)本(ben)稀(xi)釋(shi)(shi)液(ye)(ye)(ye)(ye)(ye)(ye)6mL3mL無(wu)(wu)檢測(ce)(ce)抗(kang)(kang)體(ti)(ti)(ti)-HRP10mL5mL無(wu)(wu)20×洗(xi)(xi)(xi)滌(di)(di)緩(huan)沖液(ye)(ye)(ye)(ye)(ye)(ye)25mL15mL按(an)說(shuo)(shuo)明(ming)(ming)(ming)(ming)書(shu)(shu)(shu)進(jin)行(xing)稀(xi)釋(shi)(shi)底(di)物(wu)A6mL3mL無(wu)(wu)底(di)物(wu)B6mL3mL無(wu)(wu)終(zhong)止液(ye)(ye)(ye)(ye)(ye)(ye)6mL3mL無(wu)(wu)封板(ban)(ban)(ban)膜2張(zhang)2張(zhang)無(wu)(wu)說(shuo)(shuo)明(ming)(ming)(ming)(ming)書(shu)(shu)(shu)1份1份無(wu)(wu)自(zi)封袋(dai)1個1個無(wu)(wu)注(zhu)：標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)（S0-S5）濃(nong)度(du)依(yi)次為(wei)：0、30、60、120、240、480 U/ml試(shi)(shi)(shi)劑(ji)(ji)的(de)(de)(de)(de)準(zhun)(zhun)(zhun)備(bei)(bei) 20×洗(xi)(xi)(xi)滌(di)(di)緩(huan)沖液(ye)(ye)(ye)(ye)(ye)(ye)的(de)(de)(de)(de)稀(xi)釋(shi)(shi)：蒸(zheng)餾(liu)水(shui)(shui)按(an)1：20稀(xi)釋(shi)(shi)，即(ji)1份的(de)(de)(de)(de)20×洗(xi)(xi)(xi)滌(di)(di)緩(huan)沖液(ye)(ye)(ye)(ye)(ye)(ye)加(jia)(jia)19份的(de)(de)(de)(de)蒸(zheng)餾(liu)水(shui)(shui)。洗(xi)(xi)(xi)板(ban)(ban)(ban)方法(fa)1.手工洗(xi)(xi)(xi)板(ban)(ban)(ban)：甩盡孔(kong)(kong)(kong)(kong)內(nei)(nei)液(ye)(ye)(ye)(ye)(ye)(ye)體(ti)(ti)(ti)，每孔(kong)(kong)(kong)(kong)加(jia)(jia)滿(man)洗(xi)(xi)(xi)滌(di)(di)液(ye)(ye)(ye)(ye)(ye)(ye)，靜(jing)置1min后(hou)甩盡孔(kong)(kong)(kong)(kong)內(nei)(nei)液(ye)(ye)(ye)(ye)(ye)(ye)體(ti)(ti)(ti)，在(zai)(zai)吸(xi)水(shui)(shui)紙(zhi)上(shang)拍干，如(ru)(ru)此(ci)洗(xi)(xi)(xi)板(ban)(ban)(ban)5次。2.自(zi)動洗(xi)(xi)(xi)板(ban)(ban)(ban)機：每孔(kong)(kong)(kong)(kong)注(zhu)入洗(xi)(xi)(xi)液(ye)(ye)(ye)(ye)(ye)(ye)350μL，浸泡1min，洗(xi)(xi)(xi)板(ban)(ban)(ban)5次。操(cao)(cao)作(zuo)(zuo)步驟(zou)1.從(cong)室(shi)溫(wen)(wen)平(ping)衡20min后(hou)的(de)(de)(de)(de)鋁箔袋(dai)中(zhong)(zhong)取(qu)出所需板(ban)(ban)(ban)條(tiao)，剩余(yu)板(ban)(ban)(ban)條(tiao)用(yong)自(zi)封袋(dai)密(mi)封放回4℃。2.設置標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)孔(kong)(kong)(kong)(kong)和樣(yang)(yang)本(ben)孔(kong)(kong)(kong)(kong)，標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)孔(kong)(kong)(kong)(kong)各加(jia)(jia)不(bu)(bu)同(tong)濃(nong)度(du)的(de)(de)(de)(de)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)50μL；3.樣(yang)(yang)本(ben)孔(kong)(kong)(kong)(kong)先(xian)加(jia)(jia)待測(ce)(ce)樣(yang)(yang)本(ben)10μL，再加(jia)(jia)樣(yang)(yang)本(ben)稀(xi)釋(shi)(shi)液(ye)(ye)(ye)(ye)(ye)(ye)40μL；空(kong)(kong)白(bai)孔(kong)(kong)(kong)(kong)不(bu)(bu)加(jia)(jia)。4.除(chu)(chu)空(kong)(kong)白(bai)孔(kong)(kong)(kong)(kong)外，標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)孔(kong)(kong)(kong)(kong)和樣(yang)(yang)本(ben)孔(kong)(kong)(kong)(kong)中(zhong)(zhong)每孔(kong)(kong)(kong)(kong)加(jia)(jia)入辣根過氧(yang)化(hua)物(wu)酶(mei)（HRP）標(biao)(biao)(biao)(biao)(biao)(biao)(biao)記的(de)(de)(de)(de)檢測(ce)(ce)抗(kang)(kang)體(ti)(ti)(ti)100μL，用(yong)封板(ban)(ban)(ban)膜封住反(fan)應孔(kong)(kong)(kong)(kong)，37℃水(shui)(shui)浴鍋(guo)或(huo)恒(heng)溫(wen)(wen)箱(xiang)(xiang)(xiang)溫(wen)(wen)育60min。5.棄去(qu)液(ye)(ye)(ye)(ye)(ye)(ye)體(ti)(ti)(ti)，吸(xi)水(shui)(shui)紙(zhi)上(shang)拍干，每孔(kong)(kong)(kong)(kong)加(jia)(jia)滿(man)洗(xi)(xi)(xi)滌(di)(di)液(ye)(ye)(ye)(ye)(ye)(ye)，靜(jing)置1min，甩去(qu)洗(xi)(xi)(xi)滌(di)(di)液(ye)(ye)(ye)(ye)(ye)(ye)，吸(xi)水(shui)(shui)紙(zhi)上(shang)拍干，如(ru)(ru)此(ci)重(zhong)(zhong)復洗(xi)(xi)(xi)板(ban)(ban)(ban)5次（也可(ke)(ke)用(yong)洗(xi)(xi)(xi)板(ban)(ban)(ban)機洗(xi)(xi)(xi)板(ban)(ban)(ban)）。6.每孔(kong)(kong)(kong)(kong)加(jia)(jia)入底(di)物(wu)A、B各50μL，37℃避光(guang)孵育15min。7.每孔(kong)(kong)(kong)(kong)加(jia)(jia)入終(zhong)止液(ye)(ye)(ye)(ye)(ye)(ye)50μL，15min內(nei)(nei)，在(zai)(zai)450nm波長處測(ce)(ce)定(ding)各孔(kong)(kong)(kong)(kong)的(de)(de)(de)(de)OD值(zhi)。結(jie)(jie)(jie)果(guo)判斷 繪制(zhi)(zhi)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)曲線：在(zai)(zai)Excel工作(zuo)(zuo)表中(zhong)(zhong)，以標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)濃(nong)度(du)作(zuo)(zuo)橫坐標(biao)(biao)(biao)(biao)(biao)(biao)(biao)，對應OD值(zhi)作(zuo)(zuo)縱(zong)坐標(biao)(biao)(biao)(biao)(biao)(biao)(biao)，繪制(zhi)(zhi)出標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)線性(xing)(xing)回歸曲線，按(an)曲線方程(cheng)計算各樣(yang)(yang)本(ben)濃(nong)度(du)值(zhi)。大(da)鼠纖維(wei)連(lian)(lian)接(jie)(jie)蛋(dan)白(bai)(FN) ELISA 試(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)(he) 試(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)(he)性(xing)(xing)能(neng)1.準(zhun)(zhun)(zhun)確性(xing)(xing)：標(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)線性(xing)(xing)回歸與預期濃(nong)度(du)相關系(xi)數(shu)R值(zhi)，大(da)于(yu)等于(yu)0.9900。2.靈(ling)敏度(du)：＊低(di)檢測(ce)(ce)濃(nong)度(du)小(xiao)(xiao)于(yu)1.0 U/ml。3.特異性(xing)(xing)：不(bu)(bu)與其它可(ke)(ke)溶性(xing)(xing)結(jie)(jie)(jie)構類(lei)似物(wu)交(jiao)叉反(fan)應。4.重(zhong)(zhong)復性(xing)(xing)：板(ban)(ban)(ban)內(nei)(nei)、板(ban)(ban)(ban)間變異系(xi)數(shu)均小(xiao)(xiao)于(yu)15%。5.貯藏：2-8℃，避光(guang)防潮保(bao)存。6.有效期：6個月(yue)免(mian)責聲明(ming)(ming)(ming)(ming)1.試(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)(he)僅供研(yan)究使(shi)(shi)用(yong)，不(bu)(bu)得用(yong)于(yu)臨(lin)床實(shi)驗(yan)(yan)或(huo)人體(ti)(ti)(ti)實(shi)驗(yan)(yan)，否則所產生的(de)(de)(de)(de)一(yi)切后(hou)果(guo)，由實(shi)驗(yan)(yan)者(zhe)承(cheng)擔(dan)，本(ben)公司概不(bu)(bu)負(fu)責。2.嚴格按(an)照(zhao)說(shuo)(shuo)明(ming)(ming)(ming)(ming)書(shu)(shu)(shu)操(cao)(cao)作(zuo)(zuo)，實(shi)驗(yan)(yan)者(zhe)違反(fan)說(shuo)(shuo)明(ming)(ming)(ming)(ming)書(shu)(shu)(shu)操(cao)(cao)作(zuo)(zuo)，后(hou)果(guo)由實(shi)驗(yan)(yan)者(zhe)承(cheng)擔(dan)。 大(da)鼠纖維(wei)連(lian)(lian)接(jie)(jie)蛋(dan)白(bai)(FN) ELISA 試(shi)(shi)(shi)劑(ji)(ji)盒(he)(he)(he)
大(da)(da)(da)(da)鼠(shu)乳(ru)(ru)(ru)(ru)(ru)鐵(tie)(tie)(tie)傳(chuan)遞蛋(dan)(dan)白/乳(ru)(ru)(ru)(ru)(ru)鐵(tie)(tie)(tie)蛋(dan)(dan)白(LF/LTF) ELISA 試(shi)(shi)(shi)劑(ji)(ji)(ji)(ji)(ji)(ji)盒(he)(he) 大(da)(da)(da)(da)鼠(shu)乳(ru)(ru)(ru)(ru)(ru)鐵(tie)(tie)(tie)傳(chuan)遞蛋(dan)(dan)白/乳(ru)(ru)(ru)(ru)(ru)鐵(tie)(tie)(tie)蛋(dan)(dan)白(LF/LTF) ELISA 試(shi)(shi)(shi)劑(ji)(ji)(ji)(ji)(ji)(ji)盒(he)(he) 檢(jian)測原理(li)試(shi)(shi)(shi)劑(ji)(ji)(ji)(ji)(ji)(ji)盒(he)(he)采(cai)用(yong)(yong)(yong)(yong)雙抗體(ti)(ti)(ti)(ti)(ti)一(yi)步夾心(xin)法酶(mei)聯免(mian)(mian)疫吸附試(shi)(shi)(shi)驗(yan)(yan)(yan)（ELISA）。往預先包(bao)(bao)被活(huo)(huo)性(xing)(xing)(xing)(xing)氧簇(cu)（ROS）抗體(ti)(ti)(ti)(ti)(ti)的(de)(de)(de)(de)(de)(de)(de)包(bao)(bao)被微孔(kong)(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)(zhong)，依次加入(ru)(ru)標(biao)(biao)(biao)(biao)(biao)(biao)本(ben)(ben)(ben)(ben)、標(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)、HRP標(biao)(biao)(biao)(biao)(biao)(biao)記的(de)(de)(de)(de)(de)(de)(de)檢(jian)測抗體(ti)(ti)(ti)(ti)(ti)，經過(guo)(guo)(guo)溫(wen)(wen)育并徹底洗(xi)(xi)(xi)滌(di)(di)。用(yong)(yong)(yong)(yong)底物(wu)(wu)(wu)(wu)TMB顯(xian)色，TMB在(zai)(zai)過(guo)(guo)(guo)氧化物(wu)(wu)(wu)(wu)酶(mei)的(de)(de)(de)(de)(de)(de)(de)催化下轉(zhuan)化成(cheng)藍色，并在(zai)(zai)酸的(de)(de)(de)(de)(de)(de)(de)作(zuo)(zuo)用(yong)(yong)(yong)(yong)下轉(zhuan)化成(cheng)＊終的(de)(de)(de)(de)(de)(de)(de)黃色。顏色的(de)(de)(de)(de)(de)(de)(de)深淺和(he)樣(yang)(yang)(yang)(yang)品(pin)(pin)(pin)中(zhong)(zhong)(zhong)的(de)(de)(de)(de)(de)(de)(de)活(huo)(huo)性(xing)(xing)(xing)(xing)氧簇(cu)（ROS）呈正(zheng)相關。用(yong)(yong)(yong)(yong)酶(mei)標(biao)(biao)(biao)(biao)(biao)(biao)儀(yi)在(zai)(zai)450nm 波長(chang)(chang)下測定吸光(guang)度(du)（OD 值(zhi)），計算(suan)(suan)樣(yang)(yang)(yang)(yang)品(pin)(pin)(pin)活(huo)(huo)性(xing)(xing)(xing)(xing)。大(da)(da)(da)(da)鼠(shu)乳(ru)(ru)(ru)(ru)(ru)鐵(tie)(tie)(tie)傳(chuan)遞蛋(dan)(dan)白/乳(ru)(ru)(ru)(ru)(ru)鐵(tie)(tie)(tie)蛋(dan)(dan)白(LF/LTF) ELISA 試(shi)(shi)(shi)劑(ji)(ji)(ji)(ji)(ji)(ji)盒(he)(he)樣(yang)(yang)(yang)(yang)品(pin)(pin)(pin)收集、處(chu)理(li)及(ji)保存方(fang)法1. 血清(qing)：使(shi)(shi)(shi)用(yong)(yong)(yong)(yong)不(bu)(bu)含熱原和(he)內(nei)毒素的(de)(de)(de)(de)(de)(de)(de)試(shi)(shi)(shi)管，操(cao)作(zuo)(zuo)過(guo)(guo)(guo)程中(zhong)(zhong)(zhong)避(bi)免(mian)(mian)任何(he)細(xi)胞刺激，收集血液(ye)(ye)后(hou)(hou)，3000轉(zhuan)離心(xin)10分(fen)鐘(zhong)將(jiang)血清(qing)和(he)紅細(xi)胞迅速小心(xin)地分(fen)離。2. 血漿(jiang)：EDTA、檸檬(meng)酸鹽(yan)或肝(gan)素抗凝。3000轉(zhuan)離心(xin)30分(fen)鐘(zhong)取(qu)(qu)(qu)上(shang)(shang)(shang)清(qing)。3. 細(xi)胞上(shang)(shang)(shang)清(qing)液(ye)(ye)：3000轉(zhuan)離心(xin)10分(fen)鐘(zhong)去除顆粒和(he)聚合物(wu)(wu)(wu)(wu)。4. 組織勻(yun)(yun)(yun)漿(jiang)：將(jiang)組織加入(ru)(ru)適量生(sheng)理(li)鹽(yan)水(shui)搗碎。3000轉(zhuan)離心(xin)10分(fen)鐘(zhong)取(qu)(qu)(qu)上(shang)(shang)(shang)清(qing)。5. 保存：如(ru)果(guo)樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)收集后(hou)(hou)不(bu)(bu)及(ji)時檢(jian)測，請按(an)(an)一(yi)次用(yong)(yong)(yong)(yong)量分(fen)裝，凍(dong)存于(yu)(yu)-20℃，避(bi)免(mian)(mian)反復凍(dong)融，在(zai)(zai)室(shi)(shi)(shi)溫(wen)(wen)下解凍(dong)并確(que)保樣(yang)(yang)(yang)(yang)品(pin)(pin)(pin)均(jun)(jun)勻(yun)(yun)(yun)地充分(fen)解凍(dong)。大(da)(da)(da)(da)鼠(shu)乳(ru)(ru)(ru)(ru)(ru)鐵(tie)(tie)(tie)傳(chuan)遞蛋(dan)(dan)白/乳(ru)(ru)(ru)(ru)(ru)鐵(tie)(tie)(tie)蛋(dan)(dan)白(LF/LTF) ELISA 試(shi)(shi)(shi)劑(ji)(ji)(ji)(ji)(ji)(ji)盒(he)(he) 自備物(wu)(wu)(wu)(wu)品(pin)(pin)(pin)1.酶(mei)標(biao)(biao)(biao)(biao)(biao)(biao)儀(yi)（450nm）2.高精度(du)加樣(yang)(yang)(yang)(yang)器(qi)及(ji)槍頭：0.5-10uL、2-20uL、20-200uL、200-1000uL3.37℃恒(heng)溫(wen)(wen)箱(xiang)操(cao)作(zuo)(zuo)注意(yi)事項1.試(shi)(shi)(shi)劑(ji)(ji)(ji)(ji)(ji)(ji)盒(he)(he)保存在(zai)(zai)2-8℃，使(shi)(shi)(shi)用(yong)(yong)(yong)(yong)前(qian)室(shi)(shi)(shi)溫(wen)(wen)平衡20分(fen)鐘(zhong)。從冰箱(xiang)取(qu)(qu)(qu)出的(de)(de)(de)(de)(de)(de)(de)濃縮洗(xi)(xi)(xi)滌(di)(di)液(ye)(ye)會(hui)有(you)結(jie)晶，這屬于(yu)(yu)正(zheng)常現象，水(shui)浴加熱使(shi)(shi)(shi)結(jie)晶完(wan)全(quan)溶解后(hou)(hou)再(zai)使(shi)(shi)(shi)用(yong)(yong)(yong)(yong)。2.實(shi)驗(yan)(yan)(yan)中(zhong)(zhong)(zhong)不(bu)(bu)用(yong)(yong)(yong)(yong)的(de)(de)(de)(de)(de)(de)(de)板(ban)(ban)(ban)(ban)條應(ying)立即(ji)放(fang)(fang)回(hui)(hui)自封(feng)袋(dai)中(zhong)(zhong)(zhong)，密封(feng)（低(di)(di)溫(wen)(wen)干(gan)燥(zao)）保存。3.濃度(du)為0的(de)(de)(de)(de)(de)(de)(de)S0號標(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)即(ji)可視為陰性(xing)(xing)(xing)(xing)對照或者(zhe)空(kong)白；按(an)(an)照說(shuo)明(ming)(ming)書(shu)操(cao)作(zuo)(zuo)時樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)已經稀(xi)釋(shi)(shi)5倍，＊終結(jie)果(guo)乘以5才是樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)實(shi)際濃度(du)。4.嚴格按(an)(an)照說(shuo)明(ming)(ming)書(shu)中(zhong)(zhong)(zhong)標(biao)(biao)(biao)(biao)(biao)(biao)明(ming)(ming)的(de)(de)(de)(de)(de)(de)(de)時間、加液(ye)(ye)量及(ji)順(shun)序進(jin)行溫(wen)(wen)育操(cao)作(zuo)(zuo)。5.所有(you)液(ye)(ye)體(ti)(ti)(ti)(ti)(ti)組分(fen)使(shi)(shi)(shi)用(yong)(yong)(yong)(yong)前(qian)充分(fen)搖勻(yun)(yun)(yun)。試(shi)(shi)(shi)劑(ji)(ji)(ji)(ji)(ji)(ji)盒(he)(he)組成(cheng)名稱(cheng)96孔(kong)(kong)(kong)(kong)(kong)(kong)配置48孔(kong)(kong)(kong)(kong)(kong)(kong)配置備注微孔(kong)(kong)(kong)(kong)(kong)(kong)酶(mei)標(biao)(biao)(biao)(biao)(biao)(biao)板(ban)(ban)(ban)(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)(biao)(biao)(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)0.3mL*6管0.3mL*6管無(wu)樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)稀(xi)釋(shi)(shi)液(ye)(ye)6mL3mL無(wu)檢(jian)測抗體(ti)(ti)(ti)(ti)(ti)-HRP10mL5mL無(wu)20×洗(xi)(xi)(xi)滌(di)(di)緩沖液(ye)(ye)25mL15mL按(an)(an)說(shuo)明(ming)(ming)書(shu)進(jin)行稀(xi)釋(shi)(shi)底物(wu)(wu)(wu)(wu)A6mL3mL無(wu)底物(wu)(wu)(wu)(wu)B6mL3mL無(wu)終止液(ye)(ye)6mL3mL無(wu)封(feng)板(ban)(ban)(ban)(ban)膜2張2張無(wu)說(shuo)明(ming)(ming)書(shu)1份1份無(wu)自封(feng)袋(dai)1個(ge)1個(ge)無(wu)注：標(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)（S0-S5）濃度(du)依次為：0、30、60、120、240、480 U/ml試(shi)(shi)(shi)劑(ji)(ji)(ji)(ji)(ji)(ji)的(de)(de)(de)(de)(de)(de)(de)準(zhun)(zhun)備 20×洗(xi)(xi)(xi)滌(di)(di)緩沖液(ye)(ye)的(de)(de)(de)(de)(de)(de)(de)稀(xi)釋(shi)(shi)：蒸(zheng)餾水(shui)按(an)(an)1：20稀(xi)釋(shi)(shi)，即(ji)1份的(de)(de)(de)(de)(de)(de)(de)20×洗(xi)(xi)(xi)滌(di)(di)緩沖液(ye)(ye)加19份的(de)(de)(de)(de)(de)(de)(de)蒸(zheng)餾水(shui)。洗(xi)(xi)(xi)板(ban)(ban)(ban)(ban)方(fang)法1.手工(gong)洗(xi)(xi)(xi)板(ban)(ban)(ban)(ban)：甩盡孔(kong)(kong)(kong)(kong)(kong)(kong)內(nei)液(ye)(ye)體(ti)(ti)(ti)(ti)(ti)，每孔(kong)(kong)(kong)(kong)(kong)(kong)加滿洗(xi)(xi)(xi)滌(di)(di)液(ye)(ye)，靜置1min后(hou)(hou)甩盡孔(kong)(kong)(kong)(kong)(kong)(kong)內(nei)液(ye)(ye)體(ti)(ti)(ti)(ti)(ti)，在(zai)(zai)吸水(shui)紙(zhi)上(shang)(shang)(shang)拍(pai)(pai)(pai)干(gan)，如(ru)此洗(xi)(xi)(xi)板(ban)(ban)(ban)(ban)5次。2.自動(dong)洗(xi)(xi)(xi)板(ban)(ban)(ban)(ban)機：每孔(kong)(kong)(kong)(kong)(kong)(kong)注入(ru)(ru)洗(xi)(xi)(xi)液(ye)(ye)350μL，浸泡(pao)1min，洗(xi)(xi)(xi)板(ban)(ban)(ban)(ban)5次。操(cao)作(zuo)(zuo)步驟1.從室(shi)(shi)(shi)溫(wen)(wen)平衡20min后(hou)(hou)的(de)(de)(de)(de)(de)(de)(de)鋁箔袋(dai)中(zhong)(zhong)(zhong)取(qu)(qu)(qu)出所需板(ban)(ban)(ban)(ban)條，剩余板(ban)(ban)(ban)(ban)條用(yong)(yong)(yong)(yong)自封(feng)袋(dai)密封(feng)放(fang)(fang)回(hui)(hui)4℃。2.設置標(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)品(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)(biao)準(zhun)(zhun)品(pin)(pin)(pin)孔(kong)(kong)(kong)(kong)(kong)(kong)各加不(bu)(bu)同濃度(du)的(de)(de)(de)(de)(de)(de)(de)標(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)50μL；3.樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)孔(kong)(kong)(kong)(kong)(kong)(kong)先加待(dai)測樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)10μL，再(zai)加樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)稀(xi)釋(shi)(shi)液(ye)(ye)40μL；空(kong)白孔(kong)(kong)(kong)(kong)(kong)(kong)不(bu)(bu)加。4.除空(kong)白孔(kong)(kong)(kong)(kong)(kong)(kong)外，標(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)品(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)(zhong)(zhong)每孔(kong)(kong)(kong)(kong)(kong)(kong)加入(ru)(ru)辣根過(guo)(guo)(guo)氧化物(wu)(wu)(wu)(wu)酶(mei)（HRP）標(biao)(biao)(biao)(biao)(biao)(biao)記的(de)(de)(de)(de)(de)(de)(de)檢(jian)測抗體(ti)(ti)(ti)(ti)(ti)100μL，用(yong)(yong)(yong)(yong)封(feng)板(ban)(ban)(ban)(ban)膜封(feng)住反應(ying)孔(kong)(kong)(kong)(kong)(kong)(kong)，37℃水(shui)浴鍋或恒(heng)溫(wen)(wen)箱(xiang)溫(wen)(wen)育60min。5.棄去液(ye)(ye)體(ti)(ti)(ti)(ti)(ti)，吸水(shui)紙(zhi)上(shang)(shang)(shang)拍(pai)(pai)(pai)干(gan)，每孔(kong)(kong)(kong)(kong)(kong)(kong)加滿洗(xi)(xi)(xi)滌(di)(di)液(ye)(ye)，靜置1min，甩去洗(xi)(xi)(xi)滌(di)(di)液(ye)(ye)，吸水(shui)紙(zhi)上(shang)(shang)(shang)拍(pai)(pai)(pai)干(gan)，如(ru)此重復洗(xi)(xi)(xi)板(ban)(ban)(ban)(ban)5次（也可用(yong)(yong)(yong)(yong)洗(xi)(xi)(xi)板(ban)(ban)(ban)(ban)機洗(xi)(xi)(xi)板(ban)(ban)(ban)(ban)）。6.每孔(kong)(kong)(kong)(kong)(kong)(kong)加入(ru)(ru)底物(wu)(wu)(wu)(wu)A、B各50μL，37℃避(bi)光(guang)孵育15min。7.每孔(kong)(kong)(kong)(kong)(kong)(kong)加入(ru)(ru)終止液(ye)(ye)50μL，15min內(nei)，在(zai)(zai)450nm波長(chang)(chang)處(chu)測定各孔(kong)(kong)(kong)(kong)(kong)(kong)的(de)(de)(de)(de)(de)(de)(de)OD值(zhi)。結(jie)果(guo)判斷 繪制標(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)曲線(xian)：在(zai)(zai)Excel工(gong)作(zuo)(zuo)表中(zhong)(zhong)(zhong)，以標(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)濃度(du)作(zuo)(zuo)橫(heng)坐標(biao)(biao)(biao)(biao)(biao)(biao)，對應(ying)OD值(zhi)作(zuo)(zuo)縱坐標(biao)(biao)(biao)(biao)(biao)(biao)，繪制出標(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)線(xian)性(xing)(xing)(xing)(xing)回(hui)(hui)歸(gui)曲線(xian)，按(an)(an)曲線(xian)方(fang)程計算(suan)(suan)各樣(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)濃度(du)值(zhi)。大(da)(da)(da)(da)鼠(shu)乳(ru)(ru)(ru)(ru)(ru)鐵(tie)(tie)(tie)傳(chuan)遞蛋(dan)(dan)白/乳(ru)(ru)(ru)(ru)(ru)鐵(tie)(tie)(tie)蛋(dan)(dan)白(LF/LTF) ELISA 試(shi)(shi)(shi)劑(ji)(ji)(ji)(ji)(ji)(ji)盒(he)(he) 試(shi)(shi)(shi)劑(ji)(ji)(ji)(ji)(ji)(ji)盒(he)(he)性(xing)(xing)(xing)(xing)能1.準(zhun)(zhun)確(que)性(xing)(xing)(xing)(xing)：標(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)品(pin)(pin)(pin)線(xian)性(xing)(xing)(xing)(xing)回(hui)(hui)歸(gui)與預期濃度(du)相關系(xi)數R值(zhi)，大(da)(da)(da)(da)于(yu)(yu)等于(yu)(yu)0.9900。2.靈敏度(du)：＊低(di)(di)檢(jian)測濃度(du)小于(yu)(yu)1.0 U/ml。3.特(te)異性(xing)(xing)(xing)(xing)：不(bu)(bu)與其它可溶性(xing)(xing)(xing)(xing)結(jie)構(gou)類似物(wu)(wu)(wu)(wu)交叉反應(ying)。4.重復性(xing)(xing)(xing)(xing)：板(ban)(ban)(ban)(ban)內(nei)、板(ban)(ban)(ban)(ban)間變異系(xi)數均(jun)(jun)小于(yu)(yu)15%。5.貯藏：2-8℃，避(bi)光(guang)防潮保存。6.有(you)效期：6個(ge)月(yue)免(mian)(mian)責聲明(ming)(ming)1.試(shi)(shi)(shi)劑(ji)(ji)(ji)(ji)(ji)(ji)盒(he)(he)僅(jin)供(gong)研究使(shi)(shi)(shi)用(yong)(yong)(yong)(yong)，不(bu)(bu)得用(yong)(yong)(yong)(yong)于(yu)(yu)臨床(chuang)實(shi)驗(yan)(yan)(yan)或人體(ti)(ti)(ti)(ti)(ti)實(shi)驗(yan)(yan)(yan)，否則所產生(sheng)的(de)(de)(de)(de)(de)(de)(de)一(yi)切后(hou)(hou)果(guo)，由實(shi)驗(yan)(yan)(yan)者(zhe)承(cheng)擔，本(ben)(ben)(ben)(ben)公(gong)司概不(bu)(bu)負責。2.嚴格按(an)(an)照說(shuo)明(ming)(ming)書(shu)操(cao)作(zuo)(zuo)，實(shi)驗(yan)(yan)(yan)者(zhe)違反說(shuo)明(ming)(ming)書(shu)操(cao)作(zuo)(zuo)，后(hou)(hou)果(guo)由實(shi)驗(yan)(yan)(yan)者(zhe)承(cheng)擔。 大(da)(da)(da)(da)鼠(shu)乳(ru)(ru)(ru)(ru)(ru)鐵(tie)(tie)(tie)傳(chuan)遞蛋(dan)(dan)白/乳(ru)(ru)(ru)(ru)(ru)鐵(tie)(tie)(tie)蛋(dan)(dan)白(LF/LTF) ELISA 試(shi)(shi)(shi)劑(ji)(ji)(ji)(ji)(ji)(ji)盒(he)(he)
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