Dear all,
I have some cells that I want to stain for Annexin V, but they were already fixed by PFA. Can I still do immunofluorescence on Annexin V?
Also, do I need any special buffer? As for living cells, it needs Calcium to work.
Thank you!
Hi Yimeng,
No you can not fix them cause that will affect the cell morphology and as a result phosphatydilserin protein.
Yeah you need a special buffer accompanied with the kit called binding buffer. Some suggest to use solution with enough amount of calcium of you do not have an access to binding buffer.
Best...
No, annexin V binds to phosphatidylserine which is flipped to the outer side of the cell membrane in live cells in early apoptosis. Fixed cells are dead and the compromised plasma membranes of dead cells provide a path for annexin V protein to pass through to the interior of the cell. You can stain live cells with annexin V and then fix, but not the other way around.
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