[Histonet] double staining in frozen tissue

From:Kelly Yang

Dear all,
Could someone be so kind to share or advise me on a
protocol for double staining immunofluorescence on
frozen human bladder tissue? 

1)Would it cause problem if I use FITC donkey
anti-goat IgG and Alexa 647 goat anti-rabbit together
as my secondary antibody? 
2)Which fixation is better? I had tried 4%
fornaldehyde (at room temp) for 5 min and icy acetone
(-20) for5 min, and found with acetone-fixation, the
morphology of nucleus is very poor. Any suggestion?

Any help will be highly appreciated,

Kelly Yang
Graduate student 
Department of Epidemiology
School of Public Heath
University of California, Los Angeles
ext. 57795

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