[Histonet] Re: Antigen retrieval on mouse GFAP

From:"Melissa Gonzalez"

Susan, I have to agree with Geoff on Dako's GFAP. It is fantastic and doesn't require Ag retrieval. 
It picks up astrocytes beautifully in mouse brain. I would definitely recommend DAKO if you aren't using it already. 


Melissa A. Gonzalez
Cell Genesys
500 Forbes Blvd. 
South San Francisco, CA 94080

Message: 25
Date: Fri, 23 Sep 2005 10:15:06 -0400
From: Geoff McAuliffe 
Subject: Re: [Histonet] Antigen retreival on mouse GFAP.
To: Susan.Ferrigon@sanofi-aventis.com
Cc: histonet@lists.utsouthwestern.edu
Message-ID: <43340DEA.2080803@umdnj.edu>
Content-Type: text/plain; charset=ISO-8859-1; format=flowed

Hi Susan:

    I don't know why you would need antigen retrieval for GFAP on mouse 
brain. Most people I know use DAKO's rabbit anti-cow antibody, catalogue 
# Z0334. It works very well on both fixed, frozen sections and on 
formalin-fixed wax embedded tissue. With a Vector ABC Elite kit my 
dilution for GFAP primary is 1:2000 or even more dilute, depending on 
how I intensify the DAB. I also get great results with fluorescence. 
Perhaps your trouble is somewere else?


Susan.Ferrigon@sanofi-aventis.com wrote:

>Does anyone have any experience with using trypsin as the method of antigen
>retreival when staining GFAP on mouse brains???  Any advice on
>concentration, temperature and incubation time would be appreciated.
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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029 


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