[求救] 細胞免疫螢光isotype control很明顯
最近在做細胞免疫螢光,
主要是要染膜上的蛋白質,
一抗是Mouse / IgG2b, kappa
二抗則是rabbit antimouse igG接 Alexa Fluor® 488
因此我的ISOTYPE CONTROL就選擇Mouse / IgG2b, kappa,
但是結果出來Isotype control的螢光很明顯,尤其是在核的部分,
我的步驟是先做固定,染一抗,
染完一抗再用triton 100作permeabilization,
之後以4%BSA做BLOCKING,接著染二抗約一到兩小時,
不知道是為什麼結果出來Isotype control的螢光很明顯,尤其是在核的部分,
請各位大大指導一下,
謝謝
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