; 牛(niu)(niu)表(biao)(biao)(biao)面(mian)膜免(mian)(mian)疫(yi)球(qiu)(qiu)蛋(dan)白(bai)A(mIgA) ELISA 試(shi)(shi)劑(ji)(ji)盒實(shi)驗(yan)原理(li) ELISA運用(yong)了(le)免(mian)(mian)疫(yi)學(xue)(xue)原理(li)和(he)化學(xue)(xue)反應(ying)(ying)顯色原理(li)，需(xu)要將(jiang)抗原或(huo)抗體(ti)預包被在(zai)(zai)固相載(zai)體(ti)表(biao)(biao)(biao)面(mian)，使(shi)后(hou)(hou)來(lai)形成(cheng)的抗原-抗體(ti)-酶(mei) (熒(ying)光基團(tuan)) -底物(wu)復合物(wu)黏附在(zai)(zai)載(zai)體(ti)上，通過檢(jian)測(ce)(ce)OD值(zhi)或(huo)發光值(zhi)，來(lai)檢(jian)測(ce)(ce)靶蛋(dan)白(bai)的含(han)量。 實(shi)驗(yan)所(suo)需(xu)自備(bei)物(wu)品(pin)1. 酶(mei)標(biao)(biao)(biao)儀(yi)(yi)（450nm）2. 加(jia)(jia)樣(yang)(yang)器及(ji)槍頭：0.5-10uL、2-20uL、20-200uL、200-1000uL3. 37℃恒溫(wen)(wen)箱 試(shi)(shi)劑(ji)(ji)盒組成(cheng)名稱96 孔(kong)(kong)配置(zhi)48 孔(kong)(kong)配置(zhi)備(bei)注微(wei)孔(kong)(kong)酶(mei)標(biao)(biao)(biao)板12 孔(kong)(kong)×8 條(tiao)12 孔(kong)(kong)×4 條(tiao)無(wu)(wu)(wu)標(biao)(biao)(biao)準(zhun)(zhun)品(pin)0.5mL0.5mL按說(shuo)明(ming)(ming)書(shu)(shu)進(jin)行(xing)(xing)稀(xi)釋(shi)標(biao)(biao)(biao)準(zhun)(zhun)品(pin)稀(xi)釋(shi)液(ye)(ye)(ye)6mL3mL按說(shuo)明(ming)(ming)書(shu)(shu)進(jin)行(xing)(xing)稀(xi)釋(shi)樣(yang)(yang)本(ben)稀(xi)釋(shi)液(ye)(ye)(ye)6mL3mL按說(shuo)明(ming)(ming)書(shu)(shu)進(jin)行(xing)(xing)稀(xi)釋(shi)檢(jian)測(ce)(ce)抗體(ti)-HRP6mL3mL無(wu)(wu)(wu)20×洗(xi)(xi)滌(di)(di)(di)(di)(di)緩沖(chong)液(ye)(ye)(ye)20mL20mL按說(shuo)明(ming)(ming)書(shu)(shu)進(jin)行(xing)(xing)稀(xi)釋(shi)底物(wu) A6mL3mL無(wu)(wu)(wu)底物(wu) B6mL3mL無(wu)(wu)(wu)終止液(ye)(ye)(ye)6mL3mL無(wu)(wu)(wu)封板膜2 張2 張無(wu)(wu)(wu)說(shuo)明(ming)(ming)書(shu)(shu)1 份(fen)(fen)1 份(fen)(fen)無(wu)(wu)(wu)自封袋1 個(ge)(ge)1 個(ge)(ge)無(wu)(wu)(wu) 操作(zuo)步驟1. 準(zhun)(zhun)備(bei)工(gong)作(zuo)：將(jiang)試(shi)(shi)劑(ji)(ji)盒從(cong)冰箱中(zhong)取出，室(shi)(shi)溫(wen)(wen)放置(zhi)30分(fen)鐘(zhong)，使(shi)試(shi)(shi)劑(ji)(ji)盒恢(hui)復至室(shi)(shi)溫(wen)(wen)。同時(shi)(shi)準(zhun)(zhun)備(bei)好實(shi)驗(yan)所(suo)需(xu)的材料和(he)器具。2. 加(jia)(jia)樣(yang)(yang)：在(zai)(zai)酶(mei)標(biao)(biao)(biao)板的弟(di)1個(ge)(ge)孔(kong)(kong)中(zhong)加(jia)(jia)入(ru)標(biao)(biao)(biao)準(zhun)(zhun)品(pin)50μL，然后(hou)(hou)在(zai)(zai)后(hou)(hou)續的孔(kong)(kong)中(zhong)依次加(jia)(jia)入(ru)待測(ce)(ce)樣(yang)(yang)本(ben)50μL。3. 溫(wen)(wen)育：將(jiang)酶(mei)標(biao)(biao)(biao)板密封，在(zai)(zai)37℃下溫(wen)(wen)育30分(fen)鐘(zhong)。4. 洗(xi)(xi)滌(di)(di)(di)(di)(di)：用(yong)洗(xi)(xi)滌(di)(di)(di)(di)(di)液(ye)(ye)(ye)清洗(xi)(xi)酶(mei)標(biao)(biao)(biao)板，共(gong)洗(xi)(xi)滌(di)(di)(di)(di)(di)5次。每次洗(xi)(xi)滌(di)(di)(di)(di)(di)后(hou)(hou)，用(yong)吸水紙(zhi)輕輕吸干(gan)孔(kong)(kong)內(nei)液(ye)(ye)(ye)體(ti)。5. 加(jia)(jia)酶(mei)：在(zai)(zai)每個(ge)(ge)孔(kong)(kong)中(zhong)加(jia)(jia)入(ru)酶(mei)標(biao)(biao)(biao)物(wu)100μL。6. 溫(wen)(wen)育：再次將(jiang)酶(mei)標(biao)(biao)(biao)板密封，在(zai)(zai)37℃下溫(wen)(wen)育30分(fen)鐘(zhong)。7. 洗(xi)(xi)滌(di)(di)(di)(di)(di)：用(yong)洗(xi)(xi)滌(di)(di)(di)(di)(di)液(ye)(ye)(ye)清洗(xi)(xi)酶(mei)標(biao)(biao)(biao)板，共(gong)洗(xi)(xi)滌(di)(di)(di)(di)(di)5次。每次洗(xi)(xi)滌(di)(di)(di)(di)(di)后(hou)(hou)，用(yong)吸水紙(zhi)輕輕吸干(gan)孔(kong)(kong)內(nei)液(ye)(ye)(ye)體(ti)。8. 加(jia)(jia)底物(wu)：在(zai)(zai)每個(ge)(ge)孔(kong)(kong)中(zhong)加(jia)(jia)入(ru)底物(wu)A液(ye)(ye)(ye)50μL，再加(jia)(jia)入(ru)底物(wu)B液(ye)(ye)(ye)50μL。9. 終止反應(ying)(ying)：在(zai)(zai)每個(ge)(ge)孔(kong)(kong)中(zhong)加(jia)(jia)入(ru)終止液(ye)(ye)(ye)50μL。10. 檢(jian)測(ce)(ce)：用(yong)酶(mei)標(biao)(biao)(biao)儀(yi)(yi)在(zai)(zai)450nm波(bo)長(chang)下讀取各孔(kong)(kong)的吸光度(du)(du)（OD值(zhi)）。 牛(niu)(niu)表(biao)(biao)(biao)面(mian)膜免(mian)(mian)疫(yi)球(qiu)(qiu)蛋(dan)白(bai)A(mIgA) ELISA 試(shi)(shi)劑(ji)(ji)盒結果(guo)分(fen)析根據標(biao)(biao)(biao)準(zhun)(zhun)品(pin)和(he)樣(yang)(yang)本(ben)的OD值(zhi)，繪(hui)制標(biao)(biao)(biao)準(zhun)(zhun)曲線，從(cong)而(er)計算(suan)出樣(yang)(yang)本(ben)中(zhong)角蛋(dan)白(bai)1（KRT1）的濃度(du)(du)。具體(ti)分(fen)析方法(fa)可參考(kao)試(shi)(shi)劑(ji)(ji)盒說(shuo)明(ming)(ming)書(shu)(shu)或(huo)相關文獻(xian)。注意事項1. 在(zai)(zai)操作(zuo)過程(cheng)中(zhong)，應(ying)(ying)保(bao)(bao)(bao)持酶(mei)標(biao)(biao)(biao)板的干(gan)燥，避(bi)免(mian)(mian)水分(fen)進(jin)入(ru)孔(kong)(kong)內(nei)。2. 加(jia)(jia)樣(yang)(yang)時(shi)(shi)需(xu)確保(bao)(bao)(bao)加(jia)(jia)樣(yang)(yang)準(zhun)(zhun)確、無(wu)(wu)(wu)氣泡產生(sheng)。3. 溫(wen)(wen)育和(he)洗(xi)(xi)滌(di)(di)(di)(di)(di)過程(cheng)中(zhong)需(xu)嚴(yan)格(ge)控制時(shi)(shi)間和(he)溫(wen)(wen)度(du)(du)，確保(bao)(bao)(bao)實(shi)驗(yan)結果(guo)的準(zhun)(zhun)確性。4. 在(zai)(zai)使(shi)用(yong)酶(mei)標(biao)(biao)(biao)儀(yi)(yi)讀取OD值(zhi)時(shi)(shi)，需(xu)確保(bao)(bao)(bao)波(bo)長(chang)設置(zhi)正確，避(bi)免(mian)(mian)誤差產生(sheng)。5. 在(zai)(zai)整個(ge)(ge)實(shi)驗(yan)過程(cheng)中(zhong)，需(xu)保(bao)(bao)(bao)持操作(zuo)環境的清潔和(he)整潔，避(bi)免(mian)(mian)交叉(cha)污
牛表(biao)面活(huo)性物質(zhi)關(guan)聯蛋(dan)白J(SPJ) ELISA 試(shi)劑(ji)(ji)盒(he)實驗(yan)原(yuan)(yuan)(yuan)理(li)(li) ELISA運用(yong)(yong)(yong)了免疫學原(yuan)(yuan)(yuan)理(li)(li)和化學反應顯色(se)原(yuan)(yuan)(yuan)理(li)(li)，需(xu)(xu)(xu)要將(jiang)抗(kang)原(yuan)(yuan)(yuan)或(huo)(huo)抗(kang)體(ti)(ti)預包被(bei)在(zai)(zai)(zai)固(gu)相載體(ti)(ti)表(biao)面，使(shi)后來(lai)形(xing)成的(de)(de)(de)(de)(de)抗(kang)原(yuan)(yuan)(yuan)-抗(kang)體(ti)(ti)-酶(mei)(mei)(mei)(mei) (熒光(guang)基團(tuan)) -底物復(fu)合(he)物黏(nian)附在(zai)(zai)(zai)載體(ti)(ti)上，通(tong)過(guo)檢測(ce)OD值(zhi)或(huo)(huo)發光(guang)值(zhi)，來(lai)檢測(ce)靶(ba)蛋(dan)白的(de)(de)(de)(de)(de)含(han)量。 實驗(yan)所(suo)需(xu)(xu)(xu)自備物品(pin)1. 酶(mei)(mei)(mei)(mei)標儀（450nm）2. 加(jia)樣器及槍頭(tou)：0.5-10uL、2-20uL、20-200uL、200-1000uL3. 37℃恒溫(wen)箱(xiang) 試(shi)劑(ji)(ji)盒(he)組成名稱(cheng)96 孔(kong)(kong)配置(zhi)48 孔(kong)(kong)配置(zhi)備注(zhu)微孔(kong)(kong)酶(mei)(mei)(mei)(mei)標板(ban)(ban)12 孔(kong)(kong)×8 條(tiao)12 孔(kong)(kong)×4 條(tiao)無(wu)標準(zhun)(zhun)(zhun)(zhun)品(pin)0.5mL0.5mL按說明(ming)書進行(xing)稀釋(shi)(shi)(shi)(shi)標準(zhun)(zhun)(zhun)(zhun)品(pin)稀釋(shi)(shi)(shi)(shi)液(ye)(ye)(ye)(ye)6mL3mL按說明(ming)書進行(xing)稀釋(shi)(shi)(shi)(shi)樣本(ben)稀釋(shi)(shi)(shi)(shi)液(ye)(ye)(ye)(ye)6mL3mL按說明(ming)書進行(xing)稀釋(shi)(shi)(shi)(shi)檢測(ce)抗(kang)體(ti)(ti)-HRP6mL3mL無(wu)20×洗(xi)(xi)(xi)滌(di)緩沖液(ye)(ye)(ye)(ye)20mL20mL按說明(ming)書進行(xing)稀釋(shi)(shi)(shi)(shi)底物 A6mL3mL無(wu)底物 B6mL3mL無(wu)終(zhong)(zhong)止(zhi)(zhi)液(ye)(ye)(ye)(ye)6mL3mL無(wu)封板(ban)(ban)膜2 張(zhang)2 張(zhang)無(wu)說明(ming)書1 份1 份無(wu)自封袋1 個(ge)(ge)(ge)1 個(ge)(ge)(ge)無(wu) 操作步驟1. 準(zhun)(zhun)(zhun)(zhun)備工作：將(jiang)試(shi)劑(ji)(ji)盒(he)從冰(bing)箱(xiang)中(zhong)(zhong)(zhong)取出，室溫(wen)放(fang)置(zhi)30分(fen)鐘，使(shi)試(shi)劑(ji)(ji)盒(he)恢復(fu)至室溫(wen)。同(tong)時(shi)準(zhun)(zhun)(zhun)(zhun)備好實驗(yan)所(suo)需(xu)(xu)(xu)的(de)(de)(de)(de)(de)材(cai)料和器具。2. 加(jia)樣：在(zai)(zai)(zai)酶(mei)(mei)(mei)(mei)標板(ban)(ban)的(de)(de)(de)(de)(de)弟1個(ge)(ge)(ge)孔(kong)(kong)中(zhong)(zhong)(zhong)加(jia)入(ru)(ru)(ru)標準(zhun)(zhun)(zhun)(zhun)品(pin)50μL，然后在(zai)(zai)(zai)后續的(de)(de)(de)(de)(de)孔(kong)(kong)中(zhong)(zhong)(zhong)依次(ci)加(jia)入(ru)(ru)(ru)待測(ce)樣本(ben)50μL。3. 溫(wen)育(yu)(yu)：將(jiang)酶(mei)(mei)(mei)(mei)標板(ban)(ban)密封，在(zai)(zai)(zai)37℃下溫(wen)育(yu)(yu)30分(fen)鐘。4. 洗(xi)(xi)(xi)滌(di)：用(yong)(yong)(yong)洗(xi)(xi)(xi)滌(di)液(ye)(ye)(ye)(ye)清洗(xi)(xi)(xi)酶(mei)(mei)(mei)(mei)標板(ban)(ban)，共(gong)洗(xi)(xi)(xi)滌(di)5次(ci)。每次(ci)洗(xi)(xi)(xi)滌(di)后，用(yong)(yong)(yong)吸(xi)水(shui)(shui)紙(zhi)輕輕吸(xi)干孔(kong)(kong)內(nei)液(ye)(ye)(ye)(ye)體(ti)(ti)。5. 加(jia)酶(mei)(mei)(mei)(mei)：在(zai)(zai)(zai)每個(ge)(ge)(ge)孔(kong)(kong)中(zhong)(zhong)(zhong)加(jia)入(ru)(ru)(ru)酶(mei)(mei)(mei)(mei)標物100μL。6. 溫(wen)育(yu)(yu)：再(zai)次(ci)將(jiang)酶(mei)(mei)(mei)(mei)標板(ban)(ban)密封，在(zai)(zai)(zai)37℃下溫(wen)育(yu)(yu)30分(fen)鐘。7. 洗(xi)(xi)(xi)滌(di)：用(yong)(yong)(yong)洗(xi)(xi)(xi)滌(di)液(ye)(ye)(ye)(ye)清洗(xi)(xi)(xi)酶(mei)(mei)(mei)(mei)標板(ban)(ban)，共(gong)洗(xi)(xi)(xi)滌(di)5次(ci)。每次(ci)洗(xi)(xi)(xi)滌(di)后，用(yong)(yong)(yong)吸(xi)水(shui)(shui)紙(zhi)輕輕吸(xi)干孔(kong)(kong)內(nei)液(ye)(ye)(ye)(ye)體(ti)(ti)。8. 加(jia)底物：在(zai)(zai)(zai)每個(ge)(ge)(ge)孔(kong)(kong)中(zhong)(zhong)(zhong)加(jia)入(ru)(ru)(ru)底物A液(ye)(ye)(ye)(ye)50μL，再(zai)加(jia)入(ru)(ru)(ru)底物B液(ye)(ye)(ye)(ye)50μL。9. 終(zhong)(zhong)止(zhi)(zhi)反應：在(zai)(zai)(zai)每個(ge)(ge)(ge)孔(kong)(kong)中(zhong)(zhong)(zhong)加(jia)入(ru)(ru)(ru)終(zhong)(zhong)止(zhi)(zhi)液(ye)(ye)(ye)(ye)50μL。10. 檢測(ce)：用(yong)(yong)(yong)酶(mei)(mei)(mei)(mei)標儀在(zai)(zai)(zai)450nm波長下讀取各孔(kong)(kong)的(de)(de)(de)(de)(de)吸(xi)光(guang)度(du)（OD值(zhi)）。 牛表(biao)面活(huo)性物質(zhi)關(guan)聯蛋(dan)白J(SPJ) ELISA 試(shi)劑(ji)(ji)盒(he)結(jie)果分(fen)析根據標準(zhun)(zhun)(zhun)(zhun)品(pin)和樣本(ben)的(de)(de)(de)(de)(de)OD值(zhi)，繪制標準(zhun)(zhun)(zhun)(zhun)曲線，從而計(ji)算出樣本(ben)中(zhong)(zhong)(zhong)角蛋(dan)白1（KRT1）的(de)(de)(de)(de)(de)濃度(du)。具體(ti)(ti)分(fen)析方法可(ke)參考試(shi)劑(ji)(ji)盒(he)說明(ming)書或(huo)(huo)相關(guan)文獻(xian)。注(zhu)意事項1. 在(zai)(zai)(zai)操作過(guo)程中(zhong)(zhong)(zhong)，應保(bao)(bao)(bao)(bao)持酶(mei)(mei)(mei)(mei)標板(ban)(ban)的(de)(de)(de)(de)(de)干燥，避(bi)(bi)免水(shui)(shui)分(fen)進入(ru)(ru)(ru)孔(kong)(kong)內(nei)。2. 加(jia)樣時(shi)需(xu)(xu)(xu)確保(bao)(bao)(bao)(bao)加(jia)樣準(zhun)(zhun)(zhun)(zhun)確、無(wu)氣泡產(chan)(chan)生。3. 溫(wen)育(yu)(yu)和洗(xi)(xi)(xi)滌(di)過(guo)程中(zhong)(zhong)(zhong)需(xu)(xu)(xu)嚴(yan)格控制時(shi)間和溫(wen)度(du)，確保(bao)(bao)(bao)(bao)實驗(yan)結(jie)果的(de)(de)(de)(de)(de)準(zhun)(zhun)(zhun)(zhun)確性。4. 在(zai)(zai)(zai)使(shi)用(yong)(yong)(yong)酶(mei)(mei)(mei)(mei)標儀讀取OD值(zhi)時(shi)，需(xu)(xu)(xu)確保(bao)(bao)(bao)(bao)波長設置(zhi)正確，避(bi)(bi)免誤(wu)差產(chan)(chan)生。5. 在(zai)(zai)(zai)整(zheng)個(ge)(ge)(ge)實驗(yan)過(guo)程中(zhong)(zhong)(zhong)，需(xu)(xu)(xu)保(bao)(bao)(bao)(bao)持操作環境的(de)(de)(de)(de)(de)清潔(jie)和整(zheng)潔(jie)，避(bi)(bi)免交叉污
牛(niu)羥(qian)酰谷胱甘肽水(shui)(shui)解酶(mei)(mei)(mei)(mei)(mei)(HAGH) ELISA 試(shi)(shi)劑(ji)盒(he)(he)實驗原理(li) ELISA運(yun)用(yong)(yong)(yong)了(le)免(mian)(mian)疫學(xue)原理(li)和(he)(he)(he)(he)化(hua)學(xue)反應顯色原理(li)，需(xu)(xu)要(yao)將(jiang)抗(kang)原或抗(kang)體(ti)預包被在(zai)(zai)(zai)固相(xiang)載(zai)體(ti)表(biao)面，使(shi)后(hou)(hou)來形成的(de)抗(kang)原-抗(kang)體(ti)-酶(mei)(mei)(mei)(mei)(mei) (熒(ying)光基團) -底(di)(di)物(wu)(wu)復(fu)合物(wu)(wu)黏附在(zai)(zai)(zai)載(zai)體(ti)上，通過檢測OD值(zhi)或發光值(zhi)，來檢測靶蛋(dan)(dan)白的(de)含量(liang)。 實驗所需(xu)(xu)自備(bei)(bei)物(wu)(wu)品1. 酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)(biao)儀(yi)（450nm）2. 加(jia)(jia)(jia)樣(yang)器及槍頭：0.5-10uL、2-20uL、20-200uL、200-1000uL3. 37℃恒溫(wen)箱 試(shi)(shi)劑(ji)盒(he)(he)組成名稱96 孔(kong)(kong)(kong)配(pei)置48 孔(kong)(kong)(kong)配(pei)置備(bei)(bei)注微孔(kong)(kong)(kong)酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)12 孔(kong)(kong)(kong)×8 條(tiao)(tiao)12 孔(kong)(kong)(kong)×4 條(tiao)(tiao)無(wu)標(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品0.5mL0.5mL按說(shuo)明(ming)(ming)書(shu)進行(xing)(xing)稀(xi)釋標(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品稀(xi)釋液(ye)(ye)6mL3mL按說(shuo)明(ming)(ming)書(shu)進行(xing)(xing)稀(xi)釋樣(yang)本(ben)稀(xi)釋液(ye)(ye)6mL3mL按說(shuo)明(ming)(ming)書(shu)進行(xing)(xing)稀(xi)釋檢測抗(kang)體(ti)-HRP6mL3mL無(wu)20×洗滌(di)(di)緩沖液(ye)(ye)20mL20mL按說(shuo)明(ming)(ming)書(shu)進行(xing)(xing)稀(xi)釋底(di)(di)物(wu)(wu) A6mL3mL無(wu)底(di)(di)物(wu)(wu) B6mL3mL無(wu)終(zhong)止液(ye)(ye)6mL3mL無(wu)封(feng)(feng)板(ban)膜2 張2 張無(wu)說(shuo)明(ming)(ming)書(shu)1 份1 份無(wu)自封(feng)(feng)袋1 個(ge)1 個(ge)無(wu) 操(cao)作(zuo)步驟1. 準(zhun)(zhun)(zhun)(zhun)備(bei)(bei)工(gong)作(zuo)：將(jiang)試(shi)(shi)劑(ji)盒(he)(he)從(cong)冰箱中取(qu)出，室(shi)溫(wen)放置30分(fen)鐘，使(shi)試(shi)(shi)劑(ji)盒(he)(he)恢復(fu)至室(shi)溫(wen)。同時準(zhun)(zhun)(zhun)(zhun)備(bei)(bei)好(hao)實驗所需(xu)(xu)的(de)材料和(he)(he)(he)(he)器具。2. 加(jia)(jia)(jia)樣(yang)：在(zai)(zai)(zai)酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)的(de)弟1個(ge)孔(kong)(kong)(kong)中加(jia)(jia)(jia)入(ru)(ru)標(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品50μL，然后(hou)(hou)在(zai)(zai)(zai)后(hou)(hou)續的(de)孔(kong)(kong)(kong)中依次(ci)加(jia)(jia)(jia)入(ru)(ru)待測樣(yang)本(ben)50μL。3. 溫(wen)育(yu)：將(jiang)酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)密封(feng)(feng)，在(zai)(zai)(zai)37℃下(xia)(xia)溫(wen)育(yu)30分(fen)鐘。4. 洗滌(di)(di)：用(yong)(yong)(yong)洗滌(di)(di)液(ye)(ye)清洗酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)，共洗滌(di)(di)5次(ci)。每(mei)(mei)次(ci)洗滌(di)(di)后(hou)(hou)，用(yong)(yong)(yong)吸(xi)水(shui)(shui)紙(zhi)輕輕吸(xi)干(gan)孔(kong)(kong)(kong)內液(ye)(ye)體(ti)。5. 加(jia)(jia)(jia)酶(mei)(mei)(mei)(mei)(mei)：在(zai)(zai)(zai)每(mei)(mei)個(ge)孔(kong)(kong)(kong)中加(jia)(jia)(jia)入(ru)(ru)酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)(biao)物(wu)(wu)100μL。6. 溫(wen)育(yu)：再次(ci)將(jiang)酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)密封(feng)(feng)，在(zai)(zai)(zai)37℃下(xia)(xia)溫(wen)育(yu)30分(fen)鐘。7. 洗滌(di)(di)：用(yong)(yong)(yong)洗滌(di)(di)液(ye)(ye)清洗酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)，共洗滌(di)(di)5次(ci)。每(mei)(mei)次(ci)洗滌(di)(di)后(hou)(hou)，用(yong)(yong)(yong)吸(xi)水(shui)(shui)紙(zhi)輕輕吸(xi)干(gan)孔(kong)(kong)(kong)內液(ye)(ye)體(ti)。8. 加(jia)(jia)(jia)底(di)(di)物(wu)(wu)：在(zai)(zai)(zai)每(mei)(mei)個(ge)孔(kong)(kong)(kong)中加(jia)(jia)(jia)入(ru)(ru)底(di)(di)物(wu)(wu)A液(ye)(ye)50μL，再加(jia)(jia)(jia)入(ru)(ru)底(di)(di)物(wu)(wu)B液(ye)(ye)50μL。9. 終(zhong)止反應：在(zai)(zai)(zai)每(mei)(mei)個(ge)孔(kong)(kong)(kong)中加(jia)(jia)(jia)入(ru)(ru)終(zhong)止液(ye)(ye)50μL。10. 檢測：用(yong)(yong)(yong)酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)(biao)儀(yi)在(zai)(zai)(zai)450nm波長(chang)下(xia)(xia)讀取(qu)各孔(kong)(kong)(kong)的(de)吸(xi)光度（OD值(zhi)）。 牛(niu)羥(qian)酰谷胱甘肽水(shui)(shui)解酶(mei)(mei)(mei)(mei)(mei)(HAGH) ELISA 試(shi)(shi)劑(ji)盒(he)(he)結果分(fen)析(xi)根據(ju)標(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)品和(he)(he)(he)(he)樣(yang)本(ben)的(de)OD值(zhi)，繪(hui)制標(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)曲線，從(cong)而計算出樣(yang)本(ben)中角蛋(dan)(dan)白1（KRT1）的(de)濃度。具體(ti)分(fen)析(xi)方法(fa)可(ke)參考(kao)試(shi)(shi)劑(ji)盒(he)(he)說(shuo)明(ming)(ming)書(shu)或相(xiang)關文獻。注意事項1. 在(zai)(zai)(zai)操(cao)作(zuo)過程中，應保(bao)(bao)持酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)的(de)干(gan)燥，避(bi)免(mian)(mian)水(shui)(shui)分(fen)進入(ru)(ru)孔(kong)(kong)(kong)內。2. 加(jia)(jia)(jia)樣(yang)時需(xu)(xu)確(que)保(bao)(bao)加(jia)(jia)(jia)樣(yang)準(zhun)(zhun)(zhun)(zhun)確(que)、無(wu)氣泡產(chan)生(sheng)。3. 溫(wen)育(yu)和(he)(he)(he)(he)洗滌(di)(di)過程中需(xu)(xu)嚴格控制時間和(he)(he)(he)(he)溫(wen)度，確(que)保(bao)(bao)實驗結果的(de)準(zhun)(zhun)(zhun)(zhun)確(que)性。4. 在(zai)(zai)(zai)使(shi)用(yong)(yong)(yong)酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)(biao)儀(yi)讀取(qu)OD值(zhi)時，需(xu)(xu)確(que)保(bao)(bao)波長(chang)設置正確(que)，避(bi)免(mian)(mian)誤差產(chan)生(sheng)。5. 在(zai)(zai)(zai)整個(ge)實驗過程中，需(xu)(xu)保(bao)(bao)持操(cao)作(zuo)環境的(de)清潔和(he)(he)(he)(he)整潔，避(bi)免(mian)(mian)交叉污
牛(niu)胰島素樣(yang)生長(chang)因(yin)子(zi)1受體(ti)3(IGF1R3) ELISA 試(shi)劑(ji)(ji)盒(he)(he)實驗(yan)(yan)(yan)原(yuan)(yuan)理 ELISA運用了免疫學原(yuan)(yuan)理和(he)(he)化學反應(ying)顯色原(yuan)(yuan)理，需(xu)要將抗(kang)原(yuan)(yuan)或(huo)抗(kang)體(ti)預包被(bei)在(zai)(zai)(zai)固相(xiang)(xiang)載體(ti)表面，使(shi)后來形成的抗(kang)原(yuan)(yuan)-抗(kang)體(ti)-酶(mei)(mei)(mei)(mei)(mei) (熒光基團) -底物(wu)(wu)(wu)復(fu)合物(wu)(wu)(wu)黏附在(zai)(zai)(zai)載體(ti)上，通過(guo)(guo)檢測(ce)(ce)OD值或(huo)發光值，來檢測(ce)(ce)靶(ba)蛋白(bai)的含量。 實驗(yan)(yan)(yan)所需(xu)自(zi)備(bei)物(wu)(wu)(wu)品(pin)1. 酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)(biao)儀(yi)(yi)(yi)（450nm）2. 加樣(yang)器及槍頭(tou)：0.5-10uL、2-20uL、20-200uL、200-1000uL3. 37℃恒溫(wen)箱(xiang) 試(shi)劑(ji)(ji)盒(he)(he)組成名(ming)稱96 孔(kong)(kong)配置48 孔(kong)(kong)配置備(bei)注微孔(kong)(kong)酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)(ban)12 孔(kong)(kong)×8 條12 孔(kong)(kong)×4 條無(wu)(wu)標(biao)(biao)(biao)(biao)準(zhun)品(pin)0.5mL0.5mL按(an)說(shuo)明(ming)(ming)(ming)書(shu)(shu)進(jin)行稀(xi)釋(shi)(shi)標(biao)(biao)(biao)(biao)準(zhun)品(pin)稀(xi)釋(shi)(shi)液(ye)(ye)6mL3mL按(an)說(shuo)明(ming)(ming)(ming)書(shu)(shu)進(jin)行稀(xi)釋(shi)(shi)樣(yang)本稀(xi)釋(shi)(shi)液(ye)(ye)6mL3mL按(an)說(shuo)明(ming)(ming)(ming)書(shu)(shu)進(jin)行稀(xi)釋(shi)(shi)檢測(ce)(ce)抗(kang)體(ti)-HRP6mL3mL無(wu)(wu)20×洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)緩沖(chong)液(ye)(ye)20mL20mL按(an)說(shuo)明(ming)(ming)(ming)書(shu)(shu)進(jin)行稀(xi)釋(shi)(shi)底物(wu)(wu)(wu) A6mL3mL無(wu)(wu)底物(wu)(wu)(wu) B6mL3mL無(wu)(wu)終(zhong)止液(ye)(ye)6mL3mL無(wu)(wu)封(feng)板(ban)(ban)膜2 張2 張無(wu)(wu)說(shuo)明(ming)(ming)(ming)書(shu)(shu)1 份1 份無(wu)(wu)自(zi)封(feng)袋(dai)1 個(ge)(ge)(ge)1 個(ge)(ge)(ge)無(wu)(wu) 操(cao)作步驟1. 準(zhun)備(bei)工作：將試(shi)劑(ji)(ji)盒(he)(he)從冰箱(xiang)中(zhong)(zhong)(zhong)取出，室溫(wen)放置30分(fen)鐘(zhong)(zhong)，使(shi)試(shi)劑(ji)(ji)盒(he)(he)恢復(fu)至室溫(wen)。同時準(zhun)備(bei)好實驗(yan)(yan)(yan)所需(xu)的材(cai)料和(he)(he)器具(ju)。2. 加樣(yang)：在(zai)(zai)(zai)酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)(ban)的弟1個(ge)(ge)(ge)孔(kong)(kong)中(zhong)(zhong)(zhong)加入(ru)標(biao)(biao)(biao)(biao)準(zhun)品(pin)50μL，然后在(zai)(zai)(zai)后續(xu)的孔(kong)(kong)中(zhong)(zhong)(zhong)依次加入(ru)待測(ce)(ce)樣(yang)本50μL。3. 溫(wen)育(yu)：將酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)(ban)密封(feng)，在(zai)(zai)(zai)37℃下(xia)(xia)溫(wen)育(yu)30分(fen)鐘(zhong)(zhong)。4. 洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)：用洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)液(ye)(ye)清(qing)洗(xi)(xi)(xi)(xi)(xi)酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)(ban)，共洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)5次。每(mei)次洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)后，用吸(xi)(xi)(xi)水紙(zhi)輕輕吸(xi)(xi)(xi)干(gan)孔(kong)(kong)內液(ye)(ye)體(ti)。5. 加酶(mei)(mei)(mei)(mei)(mei)：在(zai)(zai)(zai)每(mei)個(ge)(ge)(ge)孔(kong)(kong)中(zhong)(zhong)(zhong)加入(ru)酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)(biao)物(wu)(wu)(wu)100μL。6. 溫(wen)育(yu)：再次將酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)(ban)密封(feng)，在(zai)(zai)(zai)37℃下(xia)(xia)溫(wen)育(yu)30分(fen)鐘(zhong)(zhong)。7. 洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)：用洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)液(ye)(ye)清(qing)洗(xi)(xi)(xi)(xi)(xi)酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)(ban)，共洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)5次。每(mei)次洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)后，用吸(xi)(xi)(xi)水紙(zhi)輕輕吸(xi)(xi)(xi)干(gan)孔(kong)(kong)內液(ye)(ye)體(ti)。8. 加底物(wu)(wu)(wu)：在(zai)(zai)(zai)每(mei)個(ge)(ge)(ge)孔(kong)(kong)中(zhong)(zhong)(zhong)加入(ru)底物(wu)(wu)(wu)A液(ye)(ye)50μL，再加入(ru)底物(wu)(wu)(wu)B液(ye)(ye)50μL。9. 終(zhong)止反應(ying)：在(zai)(zai)(zai)每(mei)個(ge)(ge)(ge)孔(kong)(kong)中(zhong)(zhong)(zhong)加入(ru)終(zhong)止液(ye)(ye)50μL。10. 檢測(ce)(ce)：用酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)(biao)儀(yi)(yi)(yi)在(zai)(zai)(zai)450nm波長(chang)下(xia)(xia)讀取各孔(kong)(kong)的吸(xi)(xi)(xi)光度(du)(du)（OD值）。 牛(niu)胰島素樣(yang)生長(chang)因(yin)子(zi)1受體(ti)3(IGF1R3) ELISA 試(shi)劑(ji)(ji)盒(he)(he)結(jie)果分(fen)析根據標(biao)(biao)(biao)(biao)準(zhun)品(pin)和(he)(he)樣(yang)本的OD值，繪制標(biao)(biao)(biao)(biao)準(zhun)曲線，從而(er)計算出樣(yang)本中(zhong)(zhong)(zhong)角蛋白(bai)1（KRT1）的濃度(du)(du)。具(ju)體(ti)分(fen)析方法可參考試(shi)劑(ji)(ji)盒(he)(he)說(shuo)明(ming)(ming)(ming)書(shu)(shu)或(huo)相(xiang)(xiang)關文獻。注意事項1. 在(zai)(zai)(zai)操(cao)作過(guo)(guo)程中(zhong)(zhong)(zhong)，應(ying)保(bao)持(chi)(chi)酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)(biao)板(ban)(ban)的干(gan)燥，避免水分(fen)進(jin)入(ru)孔(kong)(kong)內。2. 加樣(yang)時需(xu)確(que)(que)(que)保(bao)加樣(yang)準(zhun)確(que)(que)(que)、無(wu)(wu)氣泡產生。3. 溫(wen)育(yu)和(he)(he)洗(xi)(xi)(xi)(xi)(xi)滌(di)(di)過(guo)(guo)程中(zhong)(zhong)(zhong)需(xu)嚴格控制時間和(he)(he)溫(wen)度(du)(du)，確(que)(que)(que)保(bao)實驗(yan)(yan)(yan)結(jie)果的準(zhun)確(que)(que)(que)性。4. 在(zai)(zai)(zai)使(shi)用酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)(biao)儀(yi)(yi)(yi)讀取OD值時，需(xu)確(que)(que)(que)保(bao)波長(chang)設置正確(que)(que)(que)，避免誤差(cha)產生。5. 在(zai)(zai)(zai)整(zheng)個(ge)(ge)(ge)實驗(yan)(yan)(yan)過(guo)(guo)程中(zhong)(zhong)(zhong)，需(xu)保(bao)持(chi)(chi)操(cao)作環境的清(qing)潔(jie)和(he)(he)整(zheng)潔(jie)，避免交(jiao)叉污
牛間(jian)(jian)羥腎(shen)上腺素(MN) ELISA 試劑(ji)(ji)(ji)盒(he)(he) 實(shi)(shi)驗(yan)(yan)原理(li)(li) ELISA運用(yong)(yong)(yong)了免疫學原理(li)(li)和(he)(he)化學反(fan)應(ying)顯色原理(li)(li)，需(xu)要將(jiang)抗原或(huo)抗體(ti)預包被在(zai)(zai)(zai)固相(xiang)載體(ti)表面，使(shi)后來形成的(de)(de)(de)抗原-抗體(ti)-酶(mei)(mei)(mei)(mei) (熒光(guang)(guang)基(ji)團) -底物(wu)(wu)(wu)復合物(wu)(wu)(wu)黏(nian)附在(zai)(zai)(zai)載體(ti)上，通(tong)過檢(jian)(jian)測(ce)(ce)OD值(zhi)或(huo)發(fa)光(guang)(guang)值(zhi)，來檢(jian)(jian)測(ce)(ce)靶蛋白(bai)(bai)的(de)(de)(de)含量。 實(shi)(shi)驗(yan)(yan)所(suo)需(xu)自(zi)備(bei)物(wu)(wu)(wu)品1. 酶(mei)(mei)(mei)(mei)標儀（450nm）2. 加(jia)(jia)(jia)(jia)樣(yang)(yang)器(qi)及(ji)槍頭：0.5-10uL、2-20uL、20-200uL、200-1000uL3. 37℃恒溫箱(xiang) 試劑(ji)(ji)(ji)盒(he)(he)組(zu)成名稱96 孔(kong)(kong)(kong)配(pei)置(zhi)48 孔(kong)(kong)(kong)配(pei)置(zhi)備(bei)注微(wei)孔(kong)(kong)(kong)酶(mei)(mei)(mei)(mei)標板(ban)(ban)12 孔(kong)(kong)(kong)×8 條(tiao)12 孔(kong)(kong)(kong)×4 條(tiao)無(wu)(wu)(wu)(wu)標準(zhun)品0.5mL0.5mL按(an)說(shuo)(shuo)明(ming)(ming)(ming)書(shu)(shu)進(jin)行(xing)稀釋(shi)標準(zhun)品稀釋(shi)液(ye)6mL3mL按(an)說(shuo)(shuo)明(ming)(ming)(ming)書(shu)(shu)進(jin)行(xing)稀釋(shi)樣(yang)(yang)本(ben)稀釋(shi)液(ye)6mL3mL按(an)說(shuo)(shuo)明(ming)(ming)(ming)書(shu)(shu)進(jin)行(xing)稀釋(shi)檢(jian)(jian)測(ce)(ce)抗體(ti)-HRP6mL3mL無(wu)(wu)(wu)(wu)20×洗(xi)(xi)(xi)(xi)滌(di)(di)(di)緩沖液(ye)20mL20mL按(an)說(shuo)(shuo)明(ming)(ming)(ming)書(shu)(shu)進(jin)行(xing)稀釋(shi)底物(wu)(wu)(wu) A6mL3mL無(wu)(wu)(wu)(wu)底物(wu)(wu)(wu) B6mL3mL無(wu)(wu)(wu)(wu)終止液(ye)6mL3mL無(wu)(wu)(wu)(wu)封板(ban)(ban)膜2 張2 張無(wu)(wu)(wu)(wu)說(shuo)(shuo)明(ming)(ming)(ming)書(shu)(shu)1 份(fen)1 份(fen)無(wu)(wu)(wu)(wu)自(zi)封袋(dai)1 個(ge)1 個(ge)無(wu)(wu)(wu)(wu) 操(cao)(cao)作(zuo)(zuo)步驟1. 準(zhun)備(bei)工作(zuo)(zuo)：將(jiang)試劑(ji)(ji)(ji)盒(he)(he)從冰箱(xiang)中取出，室(shi)溫放置(zhi)30分(fen)鐘(zhong)，使(shi)試劑(ji)(ji)(ji)盒(he)(he)恢復至室(shi)溫。同時準(zhun)備(bei)好實(shi)(shi)驗(yan)(yan)所(suo)需(xu)的(de)(de)(de)材料(liao)和(he)(he)器(qi)具。2. 加(jia)(jia)(jia)(jia)樣(yang)(yang)：在(zai)(zai)(zai)酶(mei)(mei)(mei)(mei)標板(ban)(ban)的(de)(de)(de)弟1個(ge)孔(kong)(kong)(kong)中加(jia)(jia)(jia)(jia)入標準(zhun)品50μL，然后在(zai)(zai)(zai)后續的(de)(de)(de)孔(kong)(kong)(kong)中依次(ci)(ci)加(jia)(jia)(jia)(jia)入待(dai)測(ce)(ce)樣(yang)(yang)本(ben)50μL。3. 溫育：將(jiang)酶(mei)(mei)(mei)(mei)標板(ban)(ban)密封，在(zai)(zai)(zai)37℃下(xia)溫育30分(fen)鐘(zhong)。4. 洗(xi)(xi)(xi)(xi)滌(di)(di)(di)：用(yong)(yong)(yong)洗(xi)(xi)(xi)(xi)滌(di)(di)(di)液(ye)清洗(xi)(xi)(xi)(xi)酶(mei)(mei)(mei)(mei)標板(ban)(ban)，共洗(xi)(xi)(xi)(xi)滌(di)(di)(di)5次(ci)(ci)。每(mei)次(ci)(ci)洗(xi)(xi)(xi)(xi)滌(di)(di)(di)后，用(yong)(yong)(yong)吸(xi)水(shui)紙輕輕吸(xi)干(gan)孔(kong)(kong)(kong)內液(ye)體(ti)。5. 加(jia)(jia)(jia)(jia)酶(mei)(mei)(mei)(mei)：在(zai)(zai)(zai)每(mei)個(ge)孔(kong)(kong)(kong)中加(jia)(jia)(jia)(jia)入酶(mei)(mei)(mei)(mei)標物(wu)(wu)(wu)100μL。6. 溫育：再次(ci)(ci)將(jiang)酶(mei)(mei)(mei)(mei)標板(ban)(ban)密封，在(zai)(zai)(zai)37℃下(xia)溫育30分(fen)鐘(zhong)。7. 洗(xi)(xi)(xi)(xi)滌(di)(di)(di)：用(yong)(yong)(yong)洗(xi)(xi)(xi)(xi)滌(di)(di)(di)液(ye)清洗(xi)(xi)(xi)(xi)酶(mei)(mei)(mei)(mei)標板(ban)(ban)，共洗(xi)(xi)(xi)(xi)滌(di)(di)(di)5次(ci)(ci)。每(mei)次(ci)(ci)洗(xi)(xi)(xi)(xi)滌(di)(di)(di)后，用(yong)(yong)(yong)吸(xi)水(shui)紙輕輕吸(xi)干(gan)孔(kong)(kong)(kong)內液(ye)體(ti)。8. 加(jia)(jia)(jia)(jia)底物(wu)(wu)(wu)：在(zai)(zai)(zai)每(mei)個(ge)孔(kong)(kong)(kong)中加(jia)(jia)(jia)(jia)入底物(wu)(wu)(wu)A液(ye)50μL，再加(jia)(jia)(jia)(jia)入底物(wu)(wu)(wu)B液(ye)50μL。9. 終止反(fan)應(ying)：在(zai)(zai)(zai)每(mei)個(ge)孔(kong)(kong)(kong)中加(jia)(jia)(jia)(jia)入終止液(ye)50μL。10. 檢(jian)(jian)測(ce)(ce)：用(yong)(yong)(yong)酶(mei)(mei)(mei)(mei)標儀在(zai)(zai)(zai)450nm波(bo)長(chang)下(xia)讀取各孔(kong)(kong)(kong)的(de)(de)(de)吸(xi)光(guang)(guang)度（OD值(zhi)）。 牛間(jian)(jian)羥腎(shen)上腺素(MN) ELISA 試劑(ji)(ji)(ji)盒(he)(he) 結(jie)果(guo)分(fen)析根(gen)據(ju)標準(zhun)品和(he)(he)樣(yang)(yang)本(ben)的(de)(de)(de)OD值(zhi)，繪制(zhi)標準(zhun)曲線，從而計算(suan)出樣(yang)(yang)本(ben)中角蛋白(bai)(bai)1（KRT1）的(de)(de)(de)濃(nong)度。具體(ti)分(fen)析方法可(ke)參考試劑(ji)(ji)(ji)盒(he)(he)說(shuo)(shuo)明(ming)(ming)(ming)書(shu)(shu)或(huo)相(xiang)關文獻。注意(yi)事項1. 在(zai)(zai)(zai)操(cao)(cao)作(zuo)(zuo)過程中，應(ying)保(bao)持(chi)酶(mei)(mei)(mei)(mei)標板(ban)(ban)的(de)(de)(de)干(gan)燥，避(bi)(bi)免水(shui)分(fen)進(jin)入孔(kong)(kong)(kong)內。2. 加(jia)(jia)(jia)(jia)樣(yang)(yang)時需(xu)確(que)保(bao)加(jia)(jia)(jia)(jia)樣(yang)(yang)準(zhun)確(que)、無(wu)(wu)(wu)(wu)氣泡產生(sheng)。3. 溫育和(he)(he)洗(xi)(xi)(xi)(xi)滌(di)(di)(di)過程中需(xu)嚴格(ge)控制(zhi)時間(jian)(jian)和(he)(he)溫度，確(que)保(bao)實(shi)(shi)驗(yan)(yan)結(jie)果(guo)的(de)(de)(de)準(zhun)確(que)性(xing)。4. 在(zai)(zai)(zai)使(shi)用(yong)(yong)(yong)酶(mei)(mei)(mei)(mei)標儀讀取OD值(zhi)時，需(xu)確(que)保(bao)波(bo)長(chang)設置(zhi)正(zheng)確(que)，避(bi)(bi)免誤差產生(sheng)。5. 在(zai)(zai)(zai)整個(ge)實(shi)(shi)驗(yan)(yan)過程中，需(xu)保(bao)持(chi)操(cao)(cao)作(zuo)(zuo)環(huan)境的(de)(de)(de)清潔(jie)和(he)(he)整潔(jie)，避(bi)(bi)免交叉污
牛質膜(mo)膜(mo)泡(pao)關(guan)聯(lian)蛋(dan)(dan)白(bai) (PLVAP) ELISA 試劑(ji)盒實(shi)驗(yan)(yan)原(yuan)理(li) ELISA運用了免(mian)疫(yi)學原(yuan)理(li)和(he)化(hua)學反應(ying)(ying)顯(xian)色原(yuan)理(li)，需(xu)要將(jiang)(jiang)抗原(yuan)或抗體(ti)預包被在(zai)(zai)(zai)固相載體(ti)表面，使(shi)后來(lai)形成(cheng)的(de)(de)(de)(de)抗原(yuan)-抗體(ti)-酶(mei)(mei)(mei)(mei)(mei) (熒(ying)光(guang)基團) -底(di)物(wu)復合(he)物(wu)黏附在(zai)(zai)(zai)載體(ti)上，通過(guo)(guo)檢(jian)測(ce)(ce)OD值(zhi)或發光(guang)值(zhi)，來(lai)檢(jian)測(ce)(ce)靶(ba)蛋(dan)(dan)白(bai)的(de)(de)(de)(de)含量。 實(shi)驗(yan)(yan)所需(xu)自(zi)備(bei)物(wu)品(pin)(pin)1. 酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)儀（450nm）2. 加(jia)(jia)樣器及槍頭(tou)：0.5-10uL、2-20uL、20-200uL、200-1000uL3. 37℃恒溫(wen)(wen)(wen)箱(xiang) 試劑(ji)盒組成(cheng)名稱96 孔(kong)(kong)(kong)配(pei)置48 孔(kong)(kong)(kong)配(pei)置備(bei)注(zhu)微孔(kong)(kong)(kong)酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)板12 孔(kong)(kong)(kong)×8 條12 孔(kong)(kong)(kong)×4 條無(wu)標(biao)(biao)(biao)準(zhun)(zhun)品(pin)(pin)0.5mL0.5mL按(an)說(shuo)(shuo)明(ming)書(shu)(shu)進(jin)行(xing)稀(xi)(xi)(xi)釋(shi)(shi)標(biao)(biao)(biao)準(zhun)(zhun)品(pin)(pin)稀(xi)(xi)(xi)釋(shi)(shi)液6mL3mL按(an)說(shuo)(shuo)明(ming)書(shu)(shu)進(jin)行(xing)稀(xi)(xi)(xi)釋(shi)(shi)樣本(ben)稀(xi)(xi)(xi)釋(shi)(shi)液6mL3mL按(an)說(shuo)(shuo)明(ming)書(shu)(shu)進(jin)行(xing)稀(xi)(xi)(xi)釋(shi)(shi)檢(jian)測(ce)(ce)抗體(ti)-HRP6mL3mL無(wu)20×洗(xi)滌(di)(di)(di)緩沖液20mL20mL按(an)說(shuo)(shuo)明(ming)書(shu)(shu)進(jin)行(xing)稀(xi)(xi)(xi)釋(shi)(shi)底(di)物(wu) A6mL3mL無(wu)底(di)物(wu) B6mL3mL無(wu)終止液6mL3mL無(wu)封(feng)(feng)板膜(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)(feng)袋(dai)1 個(ge)1 個(ge)無(wu) 操作步驟1. 準(zhun)(zhun)備(bei)工作：將(jiang)(jiang)試劑(ji)盒從冰箱(xiang)中(zhong)(zhong)(zhong)取(qu)出，室溫(wen)(wen)(wen)放置30分(fen)(fen)(fen)鐘，使(shi)試劑(ji)盒恢復至(zhi)室溫(wen)(wen)(wen)。同時準(zhun)(zhun)備(bei)好實(shi)驗(yan)(yan)所需(xu)的(de)(de)(de)(de)材料和(he)器具。2. 加(jia)(jia)樣：在(zai)(zai)(zai)酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)板的(de)(de)(de)(de)弟(di)1個(ge)孔(kong)(kong)(kong)中(zhong)(zhong)(zhong)加(jia)(jia)入(ru)標(biao)(biao)(biao)準(zhun)(zhun)品(pin)(pin)50μL，然(ran)后在(zai)(zai)(zai)后續的(de)(de)(de)(de)孔(kong)(kong)(kong)中(zhong)(zhong)(zhong)依次(ci)(ci)加(jia)(jia)入(ru)待測(ce)(ce)樣本(ben)50μL。3. 溫(wen)(wen)(wen)育(yu)：將(jiang)(jiang)酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)板密封(feng)(feng)，在(zai)(zai)(zai)37℃下溫(wen)(wen)(wen)育(yu)30分(fen)(fen)(fen)鐘。4. 洗(xi)滌(di)(di)(di)：用洗(xi)滌(di)(di)(di)液清(qing)(qing)洗(xi)酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)板，共洗(xi)滌(di)(di)(di)5次(ci)(ci)。每(mei)次(ci)(ci)洗(xi)滌(di)(di)(di)后，用吸(xi)(xi)水(shui)紙(zhi)輕輕吸(xi)(xi)干(gan)(gan)孔(kong)(kong)(kong)內(nei)(nei)(nei)液體(ti)。5. 加(jia)(jia)酶(mei)(mei)(mei)(mei)(mei)：在(zai)(zai)(zai)每(mei)個(ge)孔(kong)(kong)(kong)中(zhong)(zhong)(zhong)加(jia)(jia)入(ru)酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)物(wu)100μL。6. 溫(wen)(wen)(wen)育(yu)：再次(ci)(ci)將(jiang)(jiang)酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)板密封(feng)(feng)，在(zai)(zai)(zai)37℃下溫(wen)(wen)(wen)育(yu)30分(fen)(fen)(fen)鐘。7. 洗(xi)滌(di)(di)(di)：用洗(xi)滌(di)(di)(di)液清(qing)(qing)洗(xi)酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)板，共洗(xi)滌(di)(di)(di)5次(ci)(ci)。每(mei)次(ci)(ci)洗(xi)滌(di)(di)(di)后，用吸(xi)(xi)水(shui)紙(zhi)輕輕吸(xi)(xi)干(gan)(gan)孔(kong)(kong)(kong)內(nei)(nei)(nei)液體(ti)。8. 加(jia)(jia)底(di)物(wu)：在(zai)(zai)(zai)每(mei)個(ge)孔(kong)(kong)(kong)中(zhong)(zhong)(zhong)加(jia)(jia)入(ru)底(di)物(wu)A液50μL，再加(jia)(jia)入(ru)底(di)物(wu)B液50μL。9. 終止反應(ying)(ying)：在(zai)(zai)(zai)每(mei)個(ge)孔(kong)(kong)(kong)中(zhong)(zhong)(zhong)加(jia)(jia)入(ru)終止液50μL。10. 檢(jian)測(ce)(ce)：用酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)儀在(zai)(zai)(zai)450nm波長下讀(du)取(qu)各孔(kong)(kong)(kong)的(de)(de)(de)(de)吸(xi)(xi)光(guang)度（OD值(zhi)）。 牛質膜(mo)膜(mo)泡(pao)關(guan)聯(lian)蛋(dan)(dan)白(bai) (PLVAP) ELISA 試劑(ji)盒結(jie)果分(fen)(fen)(fen)析(xi)根據標(biao)(biao)(biao)準(zhun)(zhun)品(pin)(pin)和(he)樣本(ben)的(de)(de)(de)(de)OD值(zhi)，繪制標(biao)(biao)(biao)準(zhun)(zhun)曲線，從而計算出樣本(ben)中(zhong)(zhong)(zhong)角蛋(dan)(dan)白(bai)1（KRT1）的(de)(de)(de)(de)濃度。具體(ti)分(fen)(fen)(fen)析(xi)方法可參考試劑(ji)盒說(shuo)(shuo)明(ming)書(shu)(shu)或相關(guan)文獻(xian)。注(zhu)意事項1. 在(zai)(zai)(zai)操作過(guo)(guo)程中(zhong)(zhong)(zhong)，應(ying)(ying)保持酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)板的(de)(de)(de)(de)干(gan)(gan)燥，避免(mian)水(shui)分(fen)(fen)(fen)進(jin)入(ru)孔(kong)(kong)(kong)內(nei)(nei)(nei)。2. 加(jia)(jia)樣時需(xu)確(que)保加(jia)(jia)樣準(zhun)(zhun)確(que)、無(wu)氣泡(pao)產生。3. 溫(wen)(wen)(wen)育(yu)和(he)洗(xi)滌(di)(di)(di)過(guo)(guo)程中(zhong)(zhong)(zhong)需(xu)嚴格控制時間和(he)溫(wen)(wen)(wen)度，確(que)保實(shi)驗(yan)(yan)結(jie)果的(de)(de)(de)(de)準(zhun)(zhun)確(que)性。4. 在(zai)(zai)(zai)使(shi)用酶(mei)(mei)(mei)(mei)(mei)標(biao)(biao)(biao)儀讀(du)取(qu)OD值(zhi)時，需(xu)確(que)保波長設置正確(que)，避免(mian)誤(wu)差產生。5. 在(zai)(zai)(zai)整(zheng)個(ge)實(shi)驗(yan)(yan)過(guo)(guo)程中(zhong)(zhong)(zhong)，需(xu)保持操作環境(jing)的(de)(de)(de)(de)清(qing)(qing)潔和(he)整(zheng)潔，避免(mian)交叉(cha)污
牛信號素7A(SEMA7A) ELISA 試劑(ji)(ji)(ji)盒(he) 實驗(yan)原(yuan)理 ELISA運用(yong)(yong)(yong)了免疫學原(yuan)理和化學反應(ying)顯色原(yuan)理，需(xu)要將(jiang)抗(kang)(kang)(kang)原(yuan)或(huo)抗(kang)(kang)(kang)體預包被在(zai)(zai)(zai)(zai)(zai)固相載體表(biao)面(mian)，使(shi)后來形成的(de)抗(kang)(kang)(kang)原(yuan)-抗(kang)(kang)(kang)體-酶(mei)(mei) (熒(ying)光基團) -底物(wu)(wu)復合物(wu)(wu)黏附在(zai)(zai)(zai)(zai)(zai)載體上，通過(guo)檢測OD值(zhi)或(huo)發(fa)光值(zhi)，來檢測靶(ba)蛋(dan)白的(de)含量。 實驗(yan)所(suo)需(xu)自備(bei)物(wu)(wu)品(pin)1. 酶(mei)(mei)標(biao)(biao)(biao)(biao)(biao)(biao)儀（450nm）2. 加(jia)樣器及槍頭：0.5-10uL、2-20uL、20-200uL、200-1000uL3. 37℃恒溫(wen)箱 試劑(ji)(ji)(ji)盒(he)組(zu)成名(ming)稱(cheng)96 孔(kong)(kong)(kong)(kong)(kong)配置48 孔(kong)(kong)(kong)(kong)(kong)配置備(bei)注微孔(kong)(kong)(kong)(kong)(kong)酶(mei)(mei)標(biao)(biao)(biao)(biao)(biao)(biao)板(ban)(ban)(ban)12 孔(kong)(kong)(kong)(kong)(kong)×8 條12 孔(kong)(kong)(kong)(kong)(kong)×4 條無(wu)標(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)品(pin)0.5mL0.5mL按說明(ming)(ming)(ming)書(shu)進(jin)(jin)行(xing)稀釋(shi)(shi)標(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)品(pin)稀釋(shi)(shi)液(ye)(ye)(ye)6mL3mL按說明(ming)(ming)(ming)書(shu)進(jin)(jin)行(xing)稀釋(shi)(shi)樣本(ben)(ben)稀釋(shi)(shi)液(ye)(ye)(ye)6mL3mL按說明(ming)(ming)(ming)書(shu)進(jin)(jin)行(xing)稀釋(shi)(shi)檢測抗(kang)(kang)(kang)體-HRP6mL3mL無(wu)20×洗(xi)(xi)(xi)滌(di)(di)緩沖(chong)液(ye)(ye)(ye)20mL20mL按說明(ming)(ming)(ming)書(shu)進(jin)(jin)行(xing)稀釋(shi)(shi)底物(wu)(wu) A6mL3mL無(wu)底物(wu)(wu) B6mL3mL無(wu)終(zhong)止(zhi)液(ye)(ye)(ye)6mL3mL無(wu)封(feng)板(ban)(ban)(ban)膜(mo)2 張(zhang)2 張(zhang)無(wu)說明(ming)(ming)(ming)書(shu)1 份1 份無(wu)自封(feng)袋1 個(ge)1 個(ge)無(wu) 操作步驟1. 準(zhun)(zhun)備(bei)工作：將(jiang)試劑(ji)(ji)(ji)盒(he)從(cong)冰(bing)箱中(zhong)取(qu)出(chu)(chu)，室溫(wen)放置30分鐘(zhong)，使(shi)試劑(ji)(ji)(ji)盒(he)恢復至(zhi)室溫(wen)。同時(shi)(shi)準(zhun)(zhun)備(bei)好實驗(yan)所(suo)需(xu)的(de)材(cai)料和器具(ju)。2. 加(jia)樣：在(zai)(zai)(zai)(zai)(zai)酶(mei)(mei)標(biao)(biao)(biao)(biao)(biao)(biao)板(ban)(ban)(ban)的(de)弟1個(ge)孔(kong)(kong)(kong)(kong)(kong)中(zhong)加(jia)入標(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)品(pin)50μL，然(ran)后在(zai)(zai)(zai)(zai)(zai)后續的(de)孔(kong)(kong)(kong)(kong)(kong)中(zhong)依(yi)次(ci)(ci)加(jia)入待測樣本(ben)(ben)50μL。3. 溫(wen)育(yu)：將(jiang)酶(mei)(mei)標(biao)(biao)(biao)(biao)(biao)(biao)板(ban)(ban)(ban)密封(feng)，在(zai)(zai)(zai)(zai)(zai)37℃下(xia)溫(wen)育(yu)30分鐘(zhong)。4. 洗(xi)(xi)(xi)滌(di)(di)：用(yong)(yong)(yong)洗(xi)(xi)(xi)滌(di)(di)液(ye)(ye)(ye)清(qing)洗(xi)(xi)(xi)酶(mei)(mei)標(biao)(biao)(biao)(biao)(biao)(biao)板(ban)(ban)(ban)，共洗(xi)(xi)(xi)滌(di)(di)5次(ci)(ci)。每次(ci)(ci)洗(xi)(xi)(xi)滌(di)(di)后，用(yong)(yong)(yong)吸(xi)(xi)水(shui)紙(zhi)(zhi)輕輕吸(xi)(xi)干(gan)孔(kong)(kong)(kong)(kong)(kong)內液(ye)(ye)(ye)體。5. 加(jia)酶(mei)(mei)：在(zai)(zai)(zai)(zai)(zai)每個(ge)孔(kong)(kong)(kong)(kong)(kong)中(zhong)加(jia)入酶(mei)(mei)標(biao)(biao)(biao)(biao)(biao)(biao)物(wu)(wu)100μL。6. 溫(wen)育(yu)：再次(ci)(ci)將(jiang)酶(mei)(mei)標(biao)(biao)(biao)(biao)(biao)(biao)板(ban)(ban)(ban)密封(feng)，在(zai)(zai)(zai)(zai)(zai)37℃下(xia)溫(wen)育(yu)30分鐘(zhong)。7. 洗(xi)(xi)(xi)滌(di)(di)：用(yong)(yong)(yong)洗(xi)(xi)(xi)滌(di)(di)液(ye)(ye)(ye)清(qing)洗(xi)(xi)(xi)酶(mei)(mei)標(biao)(biao)(biao)(biao)(biao)(biao)板(ban)(ban)(ban)，共洗(xi)(xi)(xi)滌(di)(di)5次(ci)(ci)。每次(ci)(ci)洗(xi)(xi)(xi)滌(di)(di)后，用(yong)(yong)(yong)吸(xi)(xi)水(shui)紙(zhi)(zhi)輕輕吸(xi)(xi)干(gan)孔(kong)(kong)(kong)(kong)(kong)內液(ye)(ye)(ye)體。8. 加(jia)底物(wu)(wu)：在(zai)(zai)(zai)(zai)(zai)每個(ge)孔(kong)(kong)(kong)(kong)(kong)中(zhong)加(jia)入底物(wu)(wu)A液(ye)(ye)(ye)50μL，再加(jia)入底物(wu)(wu)B液(ye)(ye)(ye)50μL。9. 終(zhong)止(zhi)反應(ying)：在(zai)(zai)(zai)(zai)(zai)每個(ge)孔(kong)(kong)(kong)(kong)(kong)中(zhong)加(jia)入終(zhong)止(zhi)液(ye)(ye)(ye)50μL。10. 檢測：用(yong)(yong)(yong)酶(mei)(mei)標(biao)(biao)(biao)(biao)(biao)(biao)儀在(zai)(zai)(zai)(zai)(zai)450nm波長(chang)下(xia)讀取(qu)各孔(kong)(kong)(kong)(kong)(kong)的(de)吸(xi)(xi)光度（OD值(zhi)）。 牛信號素7A(SEMA7A) ELISA 試劑(ji)(ji)(ji)盒(he) 結果分析(xi)(xi)根(gen)據標(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)品(pin)和樣本(ben)(ben)的(de)OD值(zhi)，繪(hui)制標(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)曲(qu)線(xian)，從(cong)而計算出(chu)(chu)樣本(ben)(ben)中(zhong)角蛋(dan)白1（KRT1）的(de)濃(nong)度。具(ju)體分析(xi)(xi)方法可參考試劑(ji)(ji)(ji)盒(he)說明(ming)(ming)(ming)書(shu)或(huo)相關(guan)文獻(xian)。注意(yi)事項1. 在(zai)(zai)(zai)(zai)(zai)操作過(guo)程(cheng)中(zhong)，應(ying)保(bao)持酶(mei)(mei)標(biao)(biao)(biao)(biao)(biao)(biao)板(ban)(ban)(ban)的(de)干(gan)燥，避(bi)免水(shui)分進(jin)(jin)入孔(kong)(kong)(kong)(kong)(kong)內。2. 加(jia)樣時(shi)(shi)需(xu)確(que)(que)(que)保(bao)加(jia)樣準(zhun)(zhun)確(que)(que)(que)、無(wu)氣(qi)泡產(chan)生。3. 溫(wen)育(yu)和洗(xi)(xi)(xi)滌(di)(di)過(guo)程(cheng)中(zhong)需(xu)嚴格控制時(shi)(shi)間和溫(wen)度，確(que)(que)(que)保(bao)實驗(yan)結果的(de)準(zhun)(zhun)確(que)(que)(que)性。4. 在(zai)(zai)(zai)(zai)(zai)使(shi)用(yong)(yong)(yong)酶(mei)(mei)標(biao)(biao)(biao)(biao)(biao)(biao)儀讀取(qu)OD值(zhi)時(shi)(shi)，需(xu)確(que)(que)(que)保(bao)波長(chang)設置正確(que)(que)(que)，避(bi)免誤(wu)差產(chan)生。5. 在(zai)(zai)(zai)(zai)(zai)整個(ge)實驗(yan)過(guo)程(cheng)中(zhong)，需(xu)保(bao)持操作環(huan)境的(de)清(qing)潔和整潔，避(bi)免交叉(cha)污(wu)2
牛(niu)密封(feng)蛋(dan)白(OCLN) ELISA 試(shi)劑(ji)盒(he)(he) 實驗(yan)(yan)原理 ELISA運用(yong)了免(mian)(mian)疫學原理和化學反應(ying)顯色原理，需要將抗原或(huo)抗體(ti)(ti)(ti)預包被在(zai)固(gu)相載(zai)體(ti)(ti)(ti)表面，使(shi)(shi)后(hou)來(lai)形(xing)成(cheng)的(de)抗原-抗體(ti)(ti)(ti)-酶(mei) (熒光基團) -底物(wu)(wu)(wu)復(fu)合物(wu)(wu)(wu)黏(nian)附在(zai)載(zai)體(ti)(ti)(ti)上，通過檢測(ce)OD值(zhi)或(huo)發光值(zhi)，來(lai)檢測(ce)靶蛋(dan)白的(de)含量。 實驗(yan)(yan)所需自(zi)備物(wu)(wu)(wu)品(pin)(pin)1. 酶(mei)標(biao)(biao)(biao)(biao)(biao)儀（450nm）2. 加(jia)(jia)(jia)(jia)(jia)樣(yang)(yang)(yang)器(qi)及槍頭：0.5-10uL、2-20uL、20-200uL、200-1000uL3. 37℃恒溫(wen)(wen)箱 試(shi)劑(ji)盒(he)(he)組(zu)成(cheng)名稱96 孔(kong)(kong)(kong)配置(zhi)48 孔(kong)(kong)(kong)配置(zhi)備注微孔(kong)(kong)(kong)酶(mei)標(biao)(biao)(biao)(biao)(biao)板(ban)12 孔(kong)(kong)(kong)×8 條(tiao)12 孔(kong)(kong)(kong)×4 條(tiao)無(wu)(wu)標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)(pin)0.5mL0.5mL按(an)說(shuo)明(ming)書(shu)(shu)進行(xing)(xing)稀(xi)釋(shi)標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)(pin)稀(xi)釋(shi)液(ye)(ye)(ye)(ye)6mL3mL按(an)說(shuo)明(ming)書(shu)(shu)進行(xing)(xing)稀(xi)釋(shi)樣(yang)(yang)(yang)本稀(xi)釋(shi)液(ye)(ye)(ye)(ye)6mL3mL按(an)說(shuo)明(ming)書(shu)(shu)進行(xing)(xing)稀(xi)釋(shi)檢測(ce)抗體(ti)(ti)(ti)-HRP6mL3mL無(wu)(wu)20×洗滌(di)緩沖液(ye)(ye)(ye)(ye)20mL20mL按(an)說(shuo)明(ming)書(shu)(shu)進行(xing)(xing)稀(xi)釋(shi)底物(wu)(wu)(wu) A6mL3mL無(wu)(wu)底物(wu)(wu)(wu) B6mL3mL無(wu)(wu)終止(zhi)液(ye)(ye)(ye)(ye)6mL3mL無(wu)(wu)封(feng)板(ban)膜2 張2 張無(wu)(wu)說(shuo)明(ming)書(shu)(shu)1 份1 份無(wu)(wu)自(zi)封(feng)袋1 個1 個無(wu)(wu) 操(cao)作(zuo)步(bu)驟1. 準(zhun)(zhun)(zhun)備工作(zuo)：將試(shi)劑(ji)盒(he)(he)從冰箱中(zhong)(zhong)取(qu)出(chu)，室溫(wen)(wen)放置(zhi)30分(fen)鐘(zhong)，使(shi)(shi)試(shi)劑(ji)盒(he)(he)恢(hui)復(fu)至室溫(wen)(wen)。同時(shi)準(zhun)(zhun)(zhun)備好實驗(yan)(yan)所需的(de)材料和器(qi)具。2. 加(jia)(jia)(jia)(jia)(jia)樣(yang)(yang)(yang)：在(zai)酶(mei)標(biao)(biao)(biao)(biao)(biao)板(ban)的(de)弟1個孔(kong)(kong)(kong)中(zhong)(zhong)加(jia)(jia)(jia)(jia)(jia)入(ru)(ru)標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)(pin)50μL，然后(hou)在(zai)后(hou)續(xu)的(de)孔(kong)(kong)(kong)中(zhong)(zhong)依(yi)次(ci)加(jia)(jia)(jia)(jia)(jia)入(ru)(ru)待測(ce)樣(yang)(yang)(yang)本50μL。3. 溫(wen)(wen)育：將酶(mei)標(biao)(biao)(biao)(biao)(biao)板(ban)密封(feng)，在(zai)37℃下(xia)溫(wen)(wen)育30分(fen)鐘(zhong)。4. 洗滌(di)：用(yong)洗滌(di)液(ye)(ye)(ye)(ye)清洗酶(mei)標(biao)(biao)(biao)(biao)(biao)板(ban)，共(gong)洗滌(di)5次(ci)。每次(ci)洗滌(di)后(hou)，用(yong)吸(xi)水紙輕(qing)(qing)(qing)輕(qing)(qing)(qing)吸(xi)干(gan)孔(kong)(kong)(kong)內液(ye)(ye)(ye)(ye)體(ti)(ti)(ti)。5. 加(jia)(jia)(jia)(jia)(jia)酶(mei)：在(zai)每個孔(kong)(kong)(kong)中(zhong)(zhong)加(jia)(jia)(jia)(jia)(jia)入(ru)(ru)酶(mei)標(biao)(biao)(biao)(biao)(biao)物(wu)(wu)(wu)100μL。6. 溫(wen)(wen)育：再次(ci)將酶(mei)標(biao)(biao)(biao)(biao)(biao)板(ban)密封(feng)，在(zai)37℃下(xia)溫(wen)(wen)育30分(fen)鐘(zhong)。7. 洗滌(di)：用(yong)洗滌(di)液(ye)(ye)(ye)(ye)清洗酶(mei)標(biao)(biao)(biao)(biao)(biao)板(ban)，共(gong)洗滌(di)5次(ci)。每次(ci)洗滌(di)后(hou)，用(yong)吸(xi)水紙輕(qing)(qing)(qing)輕(qing)(qing)(qing)吸(xi)干(gan)孔(kong)(kong)(kong)內液(ye)(ye)(ye)(ye)體(ti)(ti)(ti)。8. 加(jia)(jia)(jia)(jia)(jia)底物(wu)(wu)(wu)：在(zai)每個孔(kong)(kong)(kong)中(zhong)(zhong)加(jia)(jia)(jia)(jia)(jia)入(ru)(ru)底物(wu)(wu)(wu)A液(ye)(ye)(ye)(ye)50μL，再加(jia)(jia)(jia)(jia)(jia)入(ru)(ru)底物(wu)(wu)(wu)B液(ye)(ye)(ye)(ye)50μL。9. 終止(zhi)反應(ying)：在(zai)每個孔(kong)(kong)(kong)中(zhong)(zhong)加(jia)(jia)(jia)(jia)(jia)入(ru)(ru)終止(zhi)液(ye)(ye)(ye)(ye)50μL。10. 檢測(ce)：用(yong)酶(mei)標(biao)(biao)(biao)(biao)(biao)儀在(zai)450nm波長下(xia)讀取(qu)各(ge)孔(kong)(kong)(kong)的(de)吸(xi)光度(du)(du)（OD值(zhi)）。 牛(niu)密封(feng)蛋(dan)白(OCLN) ELISA 試(shi)劑(ji)盒(he)(he)結(jie)果(guo)分(fen)析根據標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)品(pin)(pin)和樣(yang)(yang)(yang)本的(de)OD值(zhi)，繪制標(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)曲線，從而計算出(chu)樣(yang)(yang)(yang)本中(zhong)(zhong)角蛋(dan)白1（KRT1）的(de)濃度(du)(du)。具體(ti)(ti)(ti)分(fen)析方法(fa)可參考試(shi)劑(ji)盒(he)(he)說(shuo)明(ming)書(shu)(shu)或(huo)相關文獻。注意事項1. 在(zai)操(cao)作(zuo)過程中(zhong)(zhong)，應(ying)保(bao)持酶(mei)標(biao)(biao)(biao)(biao)(biao)板(ban)的(de)干(gan)燥，避免(mian)(mian)水分(fen)進入(ru)(ru)孔(kong)(kong)(kong)內。2. 加(jia)(jia)(jia)(jia)(jia)樣(yang)(yang)(yang)時(shi)需確(que)保(bao)加(jia)(jia)(jia)(jia)(jia)樣(yang)(yang)(yang)準(zhun)(zhun)(zhun)確(que)、無(wu)(wu)氣泡產生(sheng)。3. 溫(wen)(wen)育和洗滌(di)過程中(zhong)(zhong)需嚴格控(kong)制時(shi)間(jian)和溫(wen)(wen)度(du)(du)，確(que)保(bao)實驗(yan)(yan)結(jie)果(guo)的(de)準(zhun)(zhun)(zhun)確(que)性。4. 在(zai)使(shi)(shi)用(yong)酶(mei)標(biao)(biao)(biao)(biao)(biao)儀讀取(qu)OD值(zhi)時(shi)，需確(que)保(bao)波長設置(zhi)正確(que)，避免(mian)(mian)誤差產生(sheng)。5. 在(zai)整個實驗(yan)(yan)過程中(zhong)(zhong)，需保(bao)持操(cao)作(zuo)環境的(de)清潔和整潔，避免(mian)(mian)交叉(cha)污
牛丙酮酸脫氫酶(mei)(mei)β(PDHβ) ELISA 試(shi)劑(ji)盒(he)實(shi)驗(yan)原理 ELISA運用(yong)了免(mian)疫學(xue)原理和(he)(he)(he)化學(xue)反(fan)應(ying)顯色(se)原理，需(xu)(xu)要將抗原或(huo)(huo)抗體預(yu)包被在(zai)(zai)固相載體表(biao)面，使(shi)后(hou)(hou)來形成(cheng)的(de)(de)(de)抗原-抗體-酶(mei)(mei) (熒光基團(tuan)) -底(di)(di)物(wu)(wu)(wu)(wu)復合物(wu)(wu)(wu)(wu)黏(nian)附(fu)在(zai)(zai)載體上，通過(guo)檢(jian)測(ce)OD值(zhi)(zhi)或(huo)(huo)發(fa)光值(zhi)(zhi)，來檢(jian)測(ce)靶蛋(dan)白的(de)(de)(de)含量。 實(shi)驗(yan)所需(xu)(xu)自備(bei)(bei)物(wu)(wu)(wu)(wu)品(pin)1. 酶(mei)(mei)標儀（450nm）2. 加(jia)(jia)(jia)(jia)(jia)(jia)樣(yang)器及槍頭：0.5-10uL、2-20uL、20-200uL、200-1000uL3. 37℃恒溫箱 試(shi)劑(ji)盒(he)組成(cheng)名(ming)稱96 孔(kong)(kong)配(pei)置(zhi)48 孔(kong)(kong)配(pei)置(zhi)備(bei)(bei)注(zhu)微孔(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)(wu)標準品(pin)0.5mL0.5mL按說明(ming)(ming)書進(jin)(jin)行稀(xi)釋(shi)(shi)標準品(pin)稀(xi)釋(shi)(shi)液6mL3mL按說明(ming)(ming)書進(jin)(jin)行稀(xi)釋(shi)(shi)樣(yang)本(ben)(ben)稀(xi)釋(shi)(shi)液6mL3mL按說明(ming)(ming)書進(jin)(jin)行稀(xi)釋(shi)(shi)檢(jian)測(ce)抗體-HRP6mL3mL無(wu)(wu)20×洗(xi)(xi)(xi)滌(di)(di)緩沖液20mL20mL按說明(ming)(ming)書進(jin)(jin)行稀(xi)釋(shi)(shi)底(di)(di)物(wu)(wu)(wu)(wu) A6mL3mL無(wu)(wu)底(di)(di)物(wu)(wu)(wu)(wu) B6mL3mL無(wu)(wu)終止(zhi)液6mL3mL無(wu)(wu)封(feng)板(ban)(ban)膜2 張2 張無(wu)(wu)說明(ming)(ming)書1 份1 份無(wu)(wu)自封(feng)袋(dai)1 個1 個無(wu)(wu) 操(cao)作步(bu)驟1. 準備(bei)(bei)工(gong)作：將試(shi)劑(ji)盒(he)從(cong)冰(bing)箱中(zhong)(zhong)取出，室(shi)溫放置(zhi)30分(fen)(fen)鐘，使(shi)試(shi)劑(ji)盒(he)恢復至室(shi)溫。同(tong)時準備(bei)(bei)好實(shi)驗(yan)所需(xu)(xu)的(de)(de)(de)材料和(he)(he)(he)器具(ju)。2. 加(jia)(jia)(jia)(jia)(jia)(jia)樣(yang)：在(zai)(zai)酶(mei)(mei)標板(ban)(ban)的(de)(de)(de)弟(di)1個孔(kong)(kong)中(zhong)(zhong)加(jia)(jia)(jia)(jia)(jia)(jia)入(ru)(ru)(ru)標準品(pin)50μL，然后(hou)(hou)在(zai)(zai)后(hou)(hou)續的(de)(de)(de)孔(kong)(kong)中(zhong)(zhong)依次(ci)加(jia)(jia)(jia)(jia)(jia)(jia)入(ru)(ru)(ru)待測(ce)樣(yang)本(ben)(ben)50μL。3. 溫育(yu)(yu)：將酶(mei)(mei)標板(ban)(ban)密封(feng)，在(zai)(zai)37℃下(xia)溫育(yu)(yu)30分(fen)(fen)鐘。4. 洗(xi)(xi)(xi)滌(di)(di)：用(yong)洗(xi)(xi)(xi)滌(di)(di)液清(qing)洗(xi)(xi)(xi)酶(mei)(mei)標板(ban)(ban)，共(gong)(gong)洗(xi)(xi)(xi)滌(di)(di)5次(ci)。每次(ci)洗(xi)(xi)(xi)滌(di)(di)后(hou)(hou)，用(yong)吸(xi)(xi)水紙輕輕吸(xi)(xi)干(gan)孔(kong)(kong)內液體。5. 加(jia)(jia)(jia)(jia)(jia)(jia)酶(mei)(mei)：在(zai)(zai)每個孔(kong)(kong)中(zhong)(zhong)加(jia)(jia)(jia)(jia)(jia)(jia)入(ru)(ru)(ru)酶(mei)(mei)標物(wu)(wu)(wu)(wu)100μL。6. 溫育(yu)(yu)：再次(ci)將酶(mei)(mei)標板(ban)(ban)密封(feng)，在(zai)(zai)37℃下(xia)溫育(yu)(yu)30分(fen)(fen)鐘。7. 洗(xi)(xi)(xi)滌(di)(di)：用(yong)洗(xi)(xi)(xi)滌(di)(di)液清(qing)洗(xi)(xi)(xi)酶(mei)(mei)標板(ban)(ban)，共(gong)(gong)洗(xi)(xi)(xi)滌(di)(di)5次(ci)。每次(ci)洗(xi)(xi)(xi)滌(di)(di)后(hou)(hou)，用(yong)吸(xi)(xi)水紙輕輕吸(xi)(xi)干(gan)孔(kong)(kong)內液體。8. 加(jia)(jia)(jia)(jia)(jia)(jia)底(di)(di)物(wu)(wu)(wu)(wu)：在(zai)(zai)每個孔(kong)(kong)中(zhong)(zhong)加(jia)(jia)(jia)(jia)(jia)(jia)入(ru)(ru)(ru)底(di)(di)物(wu)(wu)(wu)(wu)A液50μL，再加(jia)(jia)(jia)(jia)(jia)(jia)入(ru)(ru)(ru)底(di)(di)物(wu)(wu)(wu)(wu)B液50μL。9. 終止(zhi)反(fan)應(ying)：在(zai)(zai)每個孔(kong)(kong)中(zhong)(zhong)加(jia)(jia)(jia)(jia)(jia)(jia)入(ru)(ru)(ru)終止(zhi)液50μL。10. 檢(jian)測(ce)：用(yong)酶(mei)(mei)標儀在(zai)(zai)450nm波長下(xia)讀取各孔(kong)(kong)的(de)(de)(de)吸(xi)(xi)光度（OD值(zhi)(zhi)）。 牛丙酮酸脫氫酶(mei)(mei)β(PDHβ) ELISA 試(shi)劑(ji)盒(he) 結(jie)果(guo)分(fen)(fen)析根據(ju)標準品(pin)和(he)(he)(he)樣(yang)本(ben)(ben)的(de)(de)(de)OD值(zhi)(zhi)，繪(hui)制(zhi)標準曲線(xian)，從(cong)而計(ji)算出樣(yang)本(ben)(ben)中(zhong)(zhong)角蛋(dan)白1（KRT1）的(de)(de)(de)濃度。具(ju)體分(fen)(fen)析方法可參考試(shi)劑(ji)盒(he)說明(ming)(ming)書或(huo)(huo)相關文(wen)獻。注(zhu)意事項1. 在(zai)(zai)操(cao)作過(guo)程(cheng)(cheng)中(zhong)(zhong)，應(ying)保(bao)(bao)(bao)(bao)持酶(mei)(mei)標板(ban)(ban)的(de)(de)(de)干(gan)燥，避(bi)免(mian)水分(fen)(fen)進(jin)(jin)入(ru)(ru)(ru)孔(kong)(kong)內。2. 加(jia)(jia)(jia)(jia)(jia)(jia)樣(yang)時需(xu)(xu)確(que)保(bao)(bao)(bao)(bao)加(jia)(jia)(jia)(jia)(jia)(jia)樣(yang)準確(que)、無(wu)(wu)氣泡(pao)產生。3. 溫育(yu)(yu)和(he)(he)(he)洗(xi)(xi)(xi)滌(di)(di)過(guo)程(cheng)(cheng)中(zhong)(zhong)需(xu)(xu)嚴格控制(zhi)時間(jian)和(he)(he)(he)溫度，確(que)保(bao)(bao)(bao)(bao)實(shi)驗(yan)結(jie)果(guo)的(de)(de)(de)準確(que)性。4. 在(zai)(zai)使(shi)用(yong)酶(mei)(mei)標儀讀取OD值(zhi)(zhi)時，需(xu)(xu)確(que)保(bao)(bao)(bao)(bao)波長設(she)置(zhi)正確(que)，避(bi)免(mian)誤差產生。5. 在(zai)(zai)整個實(shi)驗(yan)過(guo)程(cheng)(cheng)中(zhong)(zhong)，需(xu)(xu)保(bao)(bao)(bao)(bao)持操(cao)作環境的(de)(de)(de)清(qing)潔和(he)(he)(he)整潔，避(bi)免(mian)交叉污(wu)
牛吡哆(duo)醛激酶(mei)(mei)(PDXK) ELISA 試(shi)劑(ji)盒(he)(he)(he) 實驗(yan)原(yuan)理(li) ELISA運用(yong)(yong)了免疫學原(yuan)理(li)和(he)(he)(he)化學反應顯色原(yuan)理(li)，需(xu)(xu)要(yao)將(jiang)(jiang)抗原(yuan)或抗體(ti)預包被在(zai)(zai)固相(xiang)載體(ti)表面，使(shi)(shi)后(hou)來形成(cheng)的(de)(de)抗原(yuan)-抗體(ti)-酶(mei)(mei) (熒光基(ji)團) -底(di)物(wu)復合物(wu)黏附在(zai)(zai)載體(ti)上，通過(guo)檢測(ce)(ce)OD值(zhi)或發(fa)光值(zhi)，來檢測(ce)(ce)靶(ba)蛋(dan)白的(de)(de)含量。 實驗(yan)所需(xu)(xu)自(zi)備(bei)物(wu)品(pin)1. 酶(mei)(mei)標(biao)(biao)儀(yi)（450nm）2. 加(jia)樣(yang)器及槍頭：0.5-10uL、2-20uL、20-200uL、200-1000uL3. 37℃恒(heng)溫(wen)箱(xiang) 試(shi)劑(ji)盒(he)(he)(he)組成(cheng)名(ming)稱(cheng)96 孔(kong)(kong)配(pei)置48 孔(kong)(kong)配(pei)置備(bei)注(zhu)微孔(kong)(kong)酶(mei)(mei)標(biao)(biao)板(ban)(ban)12 孔(kong)(kong)×8 條12 孔(kong)(kong)×4 條無(wu)(wu)(wu)標(biao)(biao)準(zhun)(zhun)品(pin)0.5mL0.5mL按(an)說(shuo)(shuo)明書(shu)進(jin)行稀(xi)(xi)釋標(biao)(biao)準(zhun)(zhun)品(pin)稀(xi)(xi)釋液(ye)(ye)(ye)(ye)6mL3mL按(an)說(shuo)(shuo)明書(shu)進(jin)行稀(xi)(xi)釋樣(yang)本稀(xi)(xi)釋液(ye)(ye)(ye)(ye)6mL3mL按(an)說(shuo)(shuo)明書(shu)進(jin)行稀(xi)(xi)釋檢測(ce)(ce)抗體(ti)-HRP6mL3mL無(wu)(wu)(wu)20×洗(xi)滌(di)緩沖(chong)液(ye)(ye)(ye)(ye)20mL20mL按(an)說(shuo)(shuo)明書(shu)進(jin)行稀(xi)(xi)釋底(di)物(wu) A6mL3mL無(wu)(wu)(wu)底(di)物(wu) B6mL3mL無(wu)(wu)(wu)終(zhong)止(zhi)液(ye)(ye)(ye)(ye)6mL3mL無(wu)(wu)(wu)封(feng)板(ban)(ban)膜2 張(zhang)(zhang)2 張(zhang)(zhang)無(wu)(wu)(wu)說(shuo)(shuo)明書(shu)1 份1 份無(wu)(wu)(wu)自(zi)封(feng)袋1 個1 個無(wu)(wu)(wu) 操作步驟1. 準(zhun)(zhun)備(bei)工作：將(jiang)(jiang)試(shi)劑(ji)盒(he)(he)(he)從冰箱(xiang)中(zhong)(zhong)(zhong)取(qu)出，室溫(wen)放置30分鐘(zhong)，使(shi)(shi)試(shi)劑(ji)盒(he)(he)(he)恢復至室溫(wen)。同時準(zhun)(zhun)備(bei)好(hao)實驗(yan)所需(xu)(xu)的(de)(de)材料和(he)(he)(he)器具(ju)。2. 加(jia)樣(yang)：在(zai)(zai)酶(mei)(mei)標(biao)(biao)板(ban)(ban)的(de)(de)弟1個孔(kong)(kong)中(zhong)(zhong)(zhong)加(jia)入標(biao)(biao)準(zhun)(zhun)品(pin)50μL，然后(hou)在(zai)(zai)后(hou)續的(de)(de)孔(kong)(kong)中(zhong)(zhong)(zhong)依(yi)次(ci)(ci)(ci)加(jia)入待測(ce)(ce)樣(yang)本50μL。3. 溫(wen)育(yu)：將(jiang)(jiang)酶(mei)(mei)標(biao)(biao)板(ban)(ban)密封(feng)，在(zai)(zai)37℃下溫(wen)育(yu)30分鐘(zhong)。4. 洗(xi)滌(di)：用(yong)(yong)洗(xi)滌(di)液(ye)(ye)(ye)(ye)清洗(xi)酶(mei)(mei)標(biao)(biao)板(ban)(ban)，共(gong)洗(xi)滌(di)5次(ci)(ci)(ci)。每次(ci)(ci)(ci)洗(xi)滌(di)后(hou)，用(yong)(yong)吸水(shui)紙(zhi)輕輕吸干(gan)孔(kong)(kong)內液(ye)(ye)(ye)(ye)體(ti)。5. 加(jia)酶(mei)(mei)：在(zai)(zai)每個孔(kong)(kong)中(zhong)(zhong)(zhong)加(jia)入酶(mei)(mei)標(biao)(biao)物(wu)100μL。6. 溫(wen)育(yu)：再次(ci)(ci)(ci)將(jiang)(jiang)酶(mei)(mei)標(biao)(biao)板(ban)(ban)密封(feng)，在(zai)(zai)37℃下溫(wen)育(yu)30分鐘(zhong)。7. 洗(xi)滌(di)：用(yong)(yong)洗(xi)滌(di)液(ye)(ye)(ye)(ye)清洗(xi)酶(mei)(mei)標(biao)(biao)板(ban)(ban)，共(gong)洗(xi)滌(di)5次(ci)(ci)(ci)。每次(ci)(ci)(ci)洗(xi)滌(di)后(hou)，用(yong)(yong)吸水(shui)紙(zhi)輕輕吸干(gan)孔(kong)(kong)內液(ye)(ye)(ye)(ye)體(ti)。8. 加(jia)底(di)物(wu)：在(zai)(zai)每個孔(kong)(kong)中(zhong)(zhong)(zhong)加(jia)入底(di)物(wu)A液(ye)(ye)(ye)(ye)50μL，再加(jia)入底(di)物(wu)B液(ye)(ye)(ye)(ye)50μL。9. 終(zhong)止(zhi)反應：在(zai)(zai)每個孔(kong)(kong)中(zhong)(zhong)(zhong)加(jia)入終(zhong)止(zhi)液(ye)(ye)(ye)(ye)50μL。10. 檢測(ce)(ce)：用(yong)(yong)酶(mei)(mei)標(biao)(biao)儀(yi)在(zai)(zai)450nm波(bo)長下讀取(qu)各孔(kong)(kong)的(de)(de)吸光度（OD值(zhi)）。 牛吡哆(duo)醛激酶(mei)(mei)(PDXK) ELISA 試(shi)劑(ji)盒(he)(he)(he) 結(jie)果分析(xi)根據標(biao)(biao)準(zhun)(zhun)品(pin)和(he)(he)(he)樣(yang)本的(de)(de)OD值(zhi)，繪制標(biao)(biao)準(zhun)(zhun)曲線，從而計(ji)算出樣(yang)本中(zhong)(zhong)(zhong)角蛋(dan)白1（KRT1）的(de)(de)濃度。具(ju)體(ti)分析(xi)方法(fa)可參考試(shi)劑(ji)盒(he)(he)(he)說(shuo)(shuo)明書(shu)或相(xiang)關文獻。注(zhu)意事項1. 在(zai)(zai)操作過(guo)程中(zhong)(zhong)(zhong)，應保(bao)持酶(mei)(mei)標(biao)(biao)板(ban)(ban)的(de)(de)干(gan)燥，避(bi)免水(shui)分進(jin)入孔(kong)(kong)內。2. 加(jia)樣(yang)時需(xu)(xu)確(que)保(bao)加(jia)樣(yang)準(zhun)(zhun)確(que)、無(wu)(wu)(wu)氣泡產(chan)生。3. 溫(wen)育(yu)和(he)(he)(he)洗(xi)滌(di)過(guo)程中(zhong)(zhong)(zhong)需(xu)(xu)嚴格控制時間和(he)(he)(he)溫(wen)度，確(que)保(bao)實驗(yan)結(jie)果的(de)(de)準(zhun)(zhun)確(que)性。4. 在(zai)(zai)使(shi)(shi)用(yong)(yong)酶(mei)(mei)標(biao)(biao)儀(yi)讀取(qu)OD值(zhi)時，需(xu)(xu)確(que)保(bao)波(bo)長設置正(zheng)確(que)，避(bi)免誤差產(chan)生。5. 在(zai)(zai)整個實驗(yan)過(guo)程中(zhong)(zhong)(zhong)，需(xu)(xu)保(bao)持操作環境的(de)(de)清潔(jie)和(he)(he)(he)整潔(jie)，避(bi)免交叉(cha)污
322