I am working with MDA-MB-231 cell line. I want to check apoptosis after treating with our designed drug. I am planning to do Annexin V PI staining to check apoptosis. I have seen MDA-MB-231 rapidly get stressed and change their morphology after any kind of treatment, even after washing. May this problem result into any false positive response in presence of Annexin V? If yes what else can be an alternate procedure to check apoptosis?
Hello Kasturee,
Yes, you can definitely use Annexin/PI for cell death detection. When you are dettaching the cells, then cells are in 3D and you can't guess a real change in morphology. Since you are measuring cell death at end of your drug treatment, so drug that will not give a false positive even if their is slight change in morphology because the cells are not dead. If you handle cells delicately there will be no cell death and hence no false positive. Take care to use proper controls like unstained cells, annexin only, PI only and a dual untreated control to normalize the background fluorescence. Other methods you can use are Live PI stain, AO/EtBr (more qualitative) etc. But Annexin/PI system will also work fine with your cells.
Hope it helps.
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