Hello everyone,
I am currently using washed human platelets to stain Annexin V as a procoagulant marker. Additionally, I am staining with PerCP-CD61 to identify platelet cells. So far, I have tried using heat treatment (boiling in water for 20 minutes) and Triton to disrupt the cell membrane. Does anyone have other suggestions for the preparation of Annexin V positive control?
Thank you!
Hi Lucille,
For inducing cell death/apoptosis, I have put cells that are in a 1.5ml tube into a microwave-heated boiling water for 5min and put it back on ice for 2min, and repeat for 3 cycles, and I get ~80% annexin V + cells. This is done with mouse lymph node cells, and I am not sure if your image showed the staining for platelets in all conditions, but you don't need platelets to be your positive control for Annexin V staining.
Another way of inducing apoptosis specifically is to use a drug called staurosporine (1285, Tocris) and you can check this paper (Article The microanatomic segregation of selection by apoptosis in t...
) for how they did it in cell culture in vitro.Hope this is helpful,
Zhixin
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