Re[2]: GFP/Apoptosis

Tom Frey (Tom_Frey@bdis.com)
Wed, 30 Jul 1997 09:04:25 -0700

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?