上海现货小鼠抗卵白蛋白特异性IgE（OVE-IsgE）ELISA试剂盒齐一生物科技有限公司主要经营进口原装Elisa试剂盒，种类齐全，价格实惠，货期短。购买我公司ELISA产品客户，可享免费代测、全程技术指导：(包括售前标本收集、后期实验、实验结束后数据分析等技术问题，我公司会有专业的技术即时给您去电沟通、解决。)保证售后服务。

上海现货小鼠抗卵白蛋白特异性IgE（OVE-IsgE）ELISA试剂盒产品的用途：仅供科研究使用！
价格：全国：,到我司741653262，齐一生物科技（上海）有限公司提供大量品牌进口原装，分装，以及稳定性强的国产ELISA试剂盒，性价比*

操作步骤:
1.包被过程(注意设置空白对照,阴性对照):
将所用抗原用包被稀释液稀释到适当浓度(一般所需抗原包被量为每孔20-200μg),每孔抗原加入100μl,置37℃,4h,或4℃,24h;弃去孔中液体.(为避免蒸发,板上应加盖或将板平放在底部有湿纱布的金属湿盒中)
2.封闭酶标反应孔:
5%小牛血清置37℃封闭40min.封闭时将封闭液加满各反应孔,并去除各孔中的气泡,封闭结束后用洗涤液满孔洗涤3遍,每遍3min.
洗涤方法:吸干孔内反应液,将洗涤液注满板孔,放置2min略作摇动,吸干孔内液,倾去液体后在吸水纸上拍干
洗涤次数3次
3.加入待检测样品(建立合适的浓度梯度):
检测时一般采用1:50-1:400的稀释度,应采用较大稀释体积进行,一般保证样品吸取量>20μl.
将稀释好的样品加入酶标反应孔中,每样品少加双孔,每孔100μl,置于37℃,40-60min.用洗涤液满孔洗涤3遍,每遍3min.
4.加入酶标抗体:
酶标抗体:根据酶结合物提供商提供的参考工作稀释度进行.37℃,30-60min之间.短于30min往往结果不稳定.每孔加100μl.洗涤同前.
5.加入底物液(现用现配):
*TMB-过氧化氢尿素溶液,OPD-过氧化氢底物液系统次之.
底物加入量:每孔100μl,置37℃避光放置3-5分钟,加入终止液显色.
6.终止反应:
每孔加入终止液50μl终止反应,于20min内测定实验结果.
7.结果判断:
OPD显色后采用492nm波长,TMB反应产物检测需要450nm波长.检测时一定要首先进行空白孔系统调零.用测定标本孔的吸收值与一组阴性标本测定孔平均值的比值(P/N)表示,当P/N大于2时作为抗体的效价.(数值的大小依具体检测要求而定.)Mouse myeloperoxidase-antineutrophil cytoplasmic antibody IgG,MPO-ANCA IgG ELISA试剂盒
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