light fixation for intracellular staining

Dear Flowers, 

I was wondering if anybody has used what is described as light fixation
(0.2% PFA) followed by permeabilisation with saponin (or anything else
that has been found to be appropriate) for intracellular staining. We
are going to test some new antibodies soon, and I was thinking of
minimising the chances of conformational changes due to fixation. 

Has anybody also tried antigen retrieval on fixed cells for the same
application?

Now I'll brace myself for the out-of-office autoreplies......

 

Thank you a lot as usual,

You are a great bunch, and 

my very best wishes for a peaceful Christmas to everyone

 

 

Mara

 

 

Mara Rocchi BVM&S, PhD

Flow Cytometry Manager

Moredun Research Institute

Pentlands Science Park

Bush Loan, Penicuik

EH260PZ

Scotland, UK

 

 

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