人乙酰膽堿酯酶(AChE-Ab)抗體ELISA檢測試劑盒子科生物研發(fā)生產(chǎn),深圳子科生物ELISA試劑盒供應(yīng)商,擁有10年的研發(fā)和生產(chǎn)經(jīng)驗(yàn),擁有*的儀器設(shè)備,專業(yè)的技術(shù)人員和*的實(shí)驗(yàn)技術(shù),目前已經(jīng)建立強(qiáng)大的實(shí)驗(yàn)技術(shù)服務(wù)平臺,品牌設(shè)立為:ZIKER 。
人乙酰膽堿酯酶(AChE-Ab)抗體ELISA檢測試劑盒實(shí)驗(yàn)測定具體步驟:
1.確定本次檢測所需的已包被抗體的酶標(biāo)板孔數(shù)目,并增加 1 孔作為 TMB 空白顯色孔。 總數(shù)=樣品數(shù)+9;做雙份檢測時×2。其余重包裝好放如冰箱中。
2.將標(biāo)準(zhǔn)品各 0.1ml 依次加入一排 7 孔中,1 孔只加樣品稀釋液的作為零孔。對于血清、體液、 組織勻漿或細(xì)胞培養(yǎng)上清,直接加已用樣品稀釋液稀釋的樣品 100μ ι 。
3.酶標(biāo)板加上蓋,37℃反應(yīng) 90 分鐘。
4.反應(yīng)后用自動洗板機(jī)吸去酶標(biāo)板內(nèi)的液體;或甩去酶標(biāo)板內(nèi)液體,再對著吸水紙拍幾下。 不洗。
5.將準(zhǔn)備好的生物素抗體工作液按每孔 0.1ml 依次加入。(TMB 空白顯色孔除 外)。37℃反應(yīng) 60 分鐘。
6. 0.01M TBS 或 0.01M PBS 洗滌 3 次,每次浸泡 1 分鐘左右。
7.將準(zhǔn)備好的 ABC 工作液按每孔 0.1ml 依次加入(TMB 空白顯色孔除外)。37℃反應(yīng) 30 分鐘。
8. 0.01M TBS 或 0.01M PBS 洗滌 5 次,每次浸泡 1-2 分鐘左右。
9.按每孔 90μ ι 依次加入已在 37℃平衡 30 分鐘的 TMB 顯色液,37℃避光反應(yīng) 20-25 分鐘 (注意:顯色時間供參考,因用戶實(shí)驗(yàn)室條件差異,顯色時間會有所不同。此時肉眼可 見標(biāo)準(zhǔn)品的前 3-4 孔有明顯的梯度藍(lán)色,后 3-4 孔差別不明顯)。
10.按每孔 0.1ml 依次加入 TMB 終止液,此時藍(lán)色立轉(zhuǎn)黃色。
11.用酶標(biāo)儀在 450nm 測定 OD值。
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