免疫组化

免疫组化 Immunohistochemistry (Cytokeratin) Dewax the sections ? Rehydrate Xylene (1) 10min Xylene (2) 5min 50%Xylene+50%EtOH 10min 100%EtOH 2min 95%EtOH 1min 80%EtOH 1min 70%EtOH 1min 50%EtOH 1min PBS 2min (2N HCl，37C，30min，PBS wash) ? 00.1% Trypsin in PBS, 37C, 20min ? PBS, 5min ? 3% HO in MeOH, RT, 30min(37C，10min) 22 ? PBS, 5min*2 ? Block: 1,5% BSA, 0.5,5% NGS in PBS, RT, 1hr ? Add primary antibody (in block buffer) ? 04C, overnight ? PBS, 5min*3 ? Add secondary antibody (goat-anti-mouse HRP, in block buffer), RT, 1hr ? PBS, 5min*3 ? DAB substrate(1ml Milli-Q + 1drop A + 1drop B + 1drop C + 1drop D), RT, ,min ? Wash with tap water ? Stain with hematoxylin ? Dehydrate ? Mounting