Description
Annexin V is a phospholipid phosphatidylserine (PS) binding protein. In the earlier stages of apoptosis, PS is translocated to the outer sections of the cell plasma membrane and is exposed to the external cellular environment. The mark of the late stages of cellular death is the loss of membrane integrity. Cells can be stained with Annexin V conjugated to a fluorochrome together with 7-AAD, which is excluded from viable cells with intact membranes. This can be used to distinguish cells that are viable (Annexin V and 7-AAD negative), early apoptotic (Annexin V positive and 7-AAD negative), and those that are dead or in late apoptosis (Annexin V and 7-AAD positive). The Annexin V Apoptosis Detection Kit contains: Annexin V APC (100 tests), Annexin V Binding Buffer 1X (50mL), and 7-AAD (200 tests).
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耗材:滤纸、PVDF膜或NC膜、乳胶手套等
仪器:电泳仪、电转槽、摇床、制冰机(电转时需冰浴)等
转膜后就要区分与胶接触的一面与另一面了。话说楼主好奇的话可以跑蛋白的试试看哈。比如目的蛋白的胶用光滑面贴着胶,内参蛋白用粗糙面贴着胶一起转膜试试转膜结果看看呢。
当然做都是可以做,需要带上阴性和阳性的对照组,以区别出是抗体制备的问题还是抗原决定簇被破坏的问题。
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