A few small additions to Mario's note. First, annexin V biotin is also commercially available and indirect staining may be easier than trying to do a conjugate. Second, in my experience annexin V staining will not survive PFA fixation unless some divalent cation (calcium) is present in the fixative (and any wash steps!). I have only used bovine annexin V, but I assume that this will be true of human recombinant annexin V also. Tom Frey We haven't specifically tried GFP with annexin, but there is no reason for this not to work--we often use GFP with a wide variety of fluorochromes, like PE, Cy5PE, TR, APC, etc; annexin V can be conjugated to any of these to be used in combination with GFP. If you have the annexin V protein (it can be obtained both commercially and in collaboration with some scientists producing recombinant versions), conjugation instructions can be found at "http://cmgm.stanford.edu/~roederer/abcon/". BTW, the annexin V staining survives 0.5% paraformaldehyde fixation just fine. mr > Has anyone tried this combination? Or are there any other > apoptosis markers that would work with GFP?
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