精选优质文档-倾情为你奉上免疫组织化学染色步骤1将石蜡切片二甲苯脱蜡(二甲苯20min二甲苯10min)、入水(100%酒精5min100%酒精3min95%酒精2min80%酒精1min70%酒精1min),蒸馏水冲洗后,0.01M PBS冲洗,5min3次;2枸橼酸盐缓冲液微波抗原修复,加热使水温达92-96,维持10-15min,自然冷却至室温,0.01M PBS冲洗,5min3次;33%H2O2溶液,37,20 min,以消除内源性过氧化物酶的活性,0.01M PBS冲洗,5min3次;4正常羊血清封闭,37,30 min;5倾去多余血清,滴加一抗,372小时或者4过夜,0.01M PBS冲洗,5min3次;6滴加二抗工作液,37,30 min,0.01M PBS冲洗,5min3次;7滴加三抗工作液,37,30 min,0.01M PBS冲洗,5min3次;8DAB避光显色,显微镜下控制显色时间;90.01M PBS终止显色;10梯度酒精脱水(70%酒精1min