eGFP CD25+ mystery resolved
jenny at discordia.org
Fri Nov 4 05:12:57 EST 2005
Dear Flow people,
Thank you very much to those who responded to my question regarding
the detection of CD25+ cells on cells from eGFP mice.
Dr. Rachel Gerstein suggested initially (the first time I posted
the question) that I was having a compensation problem, or that
alternatively I could try another fluorochrome rather than anti-CD25
R-PE. Turns out that after proper compensation (following suggestions
of other individuals like Dan Rosson, Andrew Beernink and Dennis
Young.... Sorry if I forgot to mention somebody.... I feel like I am
at the Academy Awards here :)) the problem was not related to
compensation per se but to a DETECTION problem with the PE antibody.
When I switch to biotin anti-CD25 followed by a second antibody (anti-
biotin APC) I was able to detect a comparable amount of CD25+
compared to non-eGFP mice.
Thank you very much for your help,
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