As far as I know, CD1a, CD4 and CD8 work only on frozen tissue, so acetone
fixation after air drying would be the way to fix and incubate 30 min at
room temp for primary. CD30 in formalin fixed tissue needs microwave
antigen retrieval and is incubated for 2 hours at room temp. rabbit anti
human Factor XIIIa (Calbiochem La Jolla CA USA) (diluted 1:400 in PBS with
1% BSA) works well applied for 30 min at RT in formalin fixed tissue only
if preceded by a 30 min trypsin digestion treatment at 37 C . Hope this
helps.
Jeff Silverman
Southampton Hospital NY USA
peptolab@hamptons.com

----------
> From: RNAhybrid@aol.com
> To: histonet@Pathology.swmed.edu
> Subject: Questions about fixation and incubation
> Date: Saturday, January 23, 1999 1:38 PM
>
> I am a dermatologist at Mt Sinai Med Ctr in NYC.  I am workingon a
project
> where i stained for cytokines in human skin, IL-4, IL-5, IFN gamma using
> formalin fixation, overnight incubation at room temp and it worked ok.
We are
> also staining for CD-4, CD-8, CD-30, CD-1a and Factor XIIIa. Path
recommended
> we use acetone fixation and incubation overnight at 4 degrees cent. and
> results have not been so good.  I am not sure which fixation to use and
> whether to incubate at room temp or 4 deg.  Any recommendations will be
> greatly appreciated.
> Donald Rudikoff