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蝶ELISA试剂盒实验原理
本试剂盒应用双抗体夹心法测定标本中蝶水平。用纯化的蝶抗体包被微孔板,制成固相抗体,往包被单抗的微孔中依次加入蝶,再与HRP标记的蝶抗体结合,形成抗体-抗原-酶标抗体复合物,经过彻底洗涤后加底物TMB显色。TMB在HRP酶的催化下转化成蓝色,并在酸的作用下转化成最终的黄色。颜色的深浅和样品中的蝶呈正相关。用酶标仪在450nm波长下测定吸光度(OD值),通过标准曲线计算样品中蝶浓度。
试剂盒组成:
1 | 30倍浓缩洗涤液 | 20ml×1瓶 | 7 | 终止液 6ml×1瓶 |
2 | 酶标试剂 | 6ml×1瓶 | 8 | 标准品(240 ng/L) 0.5ml×1瓶 |
3 | 酶标包被板 | 12孔×8条 | 9 | 标准品稀释液 1.5ml×1瓶 |
4 | 样品稀释液 | 6ml×1瓶 | 10 | 说明书 1份 |
5 | 显色剂A液 | 6ml×1瓶 | 11 | 封板膜 2张 |
6 | 显色剂B液 | 6ml×1/瓶 | 12 | 密封袋 1个 |
蝶ELISA试剂盒ELISA试剂盒具有以下优点:
1)快速: 仅需传统夹心法ELISA 1/3的操作时间,1小时即可完成整个试验流程;
2)简单: 只需1个洗涤步骤;
3)灵活: 可单独或同时检测总蛋白及磷酸化蛋白,可在同一板上检测不同的靶蛋白;
4)灵敏: 达到甚至超过行业标准,且结果一致性良好;
5)兼容: 常规比色法检测,实验室常规酶标仪读数;
温馨提醒:ELISA加样时的防错小经验
(1)用一张写满字的纸,字越多越好,比如报纸,垫在下面,这样,加过标本的小孔看过去下面的字会变小,没加的字正常,非常容易区分。
(2)可以利用液面反光与没有加的孔加以区别
(3)在标本加完后,再把加样枪全压下,使液面产生小气泡,也可以加以区分
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