lijianyu118
发表于 2006-03-12 13:17
调整好细胞浓度后制成细胞涂片,4%多聚甲醛4oC固定过夜,0.01M PBS充分冲洗、晾干,3%H2O2室温孵育20min,0.01M PBS冲洗,滴加30μl/片羊血清封闭液,室温30min后倾去,滴加1:100稀释的一抗30μl/片(0.01M PBS稀释),置湿盒内4oC过夜,0.01M PBS洗(5min×3),滴加1:200稀释的生物素化二抗30μl/片(0.01M PBS稀释),37oC湿盒内温育30min,0.01M PBS洗(5min×3),加辣根过氧化物酶标记的链霉卵白素30μl/片,37oC孵育30min,0.01M PBS洗(5min×3),DAB显色,镜下观察显色后充分水洗,常规脱水、透明、中性树胶封片。我的实验步骤,希望有帮助。