1 原理：
将通过聚丙烯酰胺凝胶电泳分离的蛋白质转移到硝酸纤维素或PVDF膜上，然后与能特异性识别待检蛋白的抗体进行反应，洗涤去除没有结合的特异性抗体后，加入标记的、能识别特异性抗体的种属特异性抗体，反应一段时间后再次洗涤去除非特异性结合的标记抗体，加入适合标记物的检测试剂进行显色或发光等，观察有无特异性蛋白条带的出现，也可通过条带的密度大小来进行特异性蛋白的半定量。
2 操作过程
SDS-PAGE电泳→转膜（PVDF或硝酸纤维素膜）
封闭→一抗→洗涤→酶标二抗反应
洗涤→显色或化学发光显影
Western blot analysis of the cleavage of Caspase-3. IM9/Bcl-2 Cells were treated with 20mM of gossypol for 4, 8
and 16 h. After treatment, cells were harvested and lysed in lysis buffer. 50ug of protein was loaded in each lane and the expression of actin was detected as a loading control.
Cytochrome c release from mitochondria to cytosol in gossypol-treated IM-9/Bcl-2 cells. Cells were treated with 10μM gossypol for different times, cytosol and mitochondrial protein were subject to SDS-PAGE followed by immunoblot with cytochrome c specific antibody.
