eGFP CD25+ mystery resolved

Jenny Zilberberg 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,
JZ



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