Problem with phalloidin staining - (Aug/10/2005 )

Hi !

I'm trying to do some actin immunofluorescence on cells using phalloidin. I've bought Alexa 488 phalloidin from Molecular Probes and followed exactly their protocol but it doesn't work.
Do you have any experience with it ? good tricks, advice ?
Thanks very much for your help.

Anne

Could you be over fixing or exposing the sample to light?

Hey Anne,
triton permeabilization appears to be crucial...0.1% 5 minsx3 times
works good