ELISA試劑盒中的酶標(biāo)用于檢測未知抗原的雙抗體夾心法:
1.包被用0.05M PH9.牰碳酸鹽包被緩沖液將抗體稀釋至蛋白質(zhì)含量為110μgml。
在每個聚苯乙烯板的反應(yīng)孔中加0.1ml4℃過夜。次日棄去孔內(nèi)溶液用洗滌緩沖液洗3次每次3分鐘。(簡稱洗滌下同)。
2.加樣加一定稀釋的待檢樣品0.1ml于上述已包被之反應(yīng)孔中置37℃孵育1小時。然后洗滌。(同時做空白孔陰性對照孔及陽性對照孔)。
3.加酶標(biāo)抗體于各反應(yīng)孔中加入新鮮稀釋的酶標(biāo)抗體(經(jīng)滴定后的稀釋度)0.1ml。37℃孵育0.51小時洗滌。
4.加底物液顯色于各反應(yīng)孔中加入臨時配制的TMB底物溶液0.1ml37℃1030分鐘。
5.終止反應(yīng)于各反應(yīng)孔中加入2M硫酸0.05ml。
6.結(jié)果判定可于白色背景上,直接用肉眼觀察結(jié)果,反應(yīng)孔內(nèi)顏色越深,陽性程度越強(qiáng);陰性反應(yīng)為無色或極淺。
依據(jù)ELISA試劑盒檢測所呈顏色的深淺以“+”、“-”號表示。也可測O·D值在ELX808酶標(biāo)儀上于450nm(若以ABTS顯色則410nm)處以空白對照孔調(diào)零后測各孔O·D值若大于規(guī)定的陰性對照OD值的2.1倍即為陽性。
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© 2015 USCAP, Inc All rights reserved 0023-6837/15
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使用產(chǎn)品:α葡萄糖苷酶ELISA試劑盒