RE: Questions about fixation and incubation

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From:"Hewlett Bryan (CMH)" <HEWLETT@EXCHANGE1.CMH.ON.CA> (by way of histonet)
To:histonet <>
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Antibodies against CD1a, CD4 and CD8 are available for formalin fixed
paraffin sections. They work well! I believe Donald was referring to
formalin fixed, frozen sections for cytokines and staining for CD's in
addition! I see no reason to suspect that Mab's that work in paraffin
would not work on FF-frozens also.


>From: 	Jeff Silverman[]
>Sent: 	January 23, 1999 5:55 PM
>Subject: 	Re: Questions about fixation and incubation
>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
>Jeff Silverman
>Southampton Hospital NY USA
>> From:
>> To:
>> 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
>> 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
>> 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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