Re: Questions about fixation and incubation

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From:Jamie Erickson <> (by way of histonet)
To:histonet <>
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Hi Donald,
              I'm very interested in your work. I'm trying to stain w/ a
human antibody  (IFN-g) in monkey skin  biopsies treated with a
inflammatory agent. I use Acetone fixed frozen sections then post fix with
formalin prior to staining. I'm also trying to get CD4/8 to work in theses
samples that are paraffin embedded. The protocol I use from R&D systems and
It incubates overnight at room temperature. If you got this to stain w/ DAB
or Fitc I'd like to know exactly what you did. Please respond, you can also
reach me  by phone at 978-623-1348.

Jamie Erickson
Genetics Institute
Andover, MA

>>> <> 01/23 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


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