Dear Flowers: Help!!!!!!! Are they any specific tricks to detecting signalling proteins like p53, NFkappab etc by flow. I have mouse lymphs induced invitro (4-18h in culture with the inducing agent) to undergo apoptosis. Then stained for surface CD45 followed by fix/perm and staining for intracellular Caspase 3 and/or said signalling molecule. I use a FITC rabbit anti mouse p53 from Caltag and PE Caspase 3 (rabbit polyclonal) from BD. The Cas3 works fine but I dont see any p53 on flow. I can detect p53 on western blots so it is there for sure. Any comments, suggestions.............??? Yasmin
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