[Histonet] GFP and B-Gal positive control slides

Gayle Callis gcallis <@t> montana.edu
Fri Dec 8 11:59:35 CST 2006


A question: How effective is your antibody without having to use TSA 

>The GFP antibodies we use are quirky so I don't want to make a
>recommendation.  At Jax where you can get the Tie-2/lacZ transgenic mice,
>you can also get GFP expressing mice for controls.  I'm not recommending,
>just a starting point for your search.  Look under research models on their
>web site.
>For the Tie-2/lac Z mice.  Fix in formalin and process to paraffin as usual.
>We use an antibody from Cortex Biochem (San Leandro, CA) that is IgG
>fraction on rabbit polyclonal to beta-galactosidase.  Around 2-3 ug/ml,
>after a 5 min proteinase K digestion.  We use an amplification kit of the
>third step (TSA for 5 minutes).  DAB, etc.  The signal is outstanding.
>Raymond Koelling
>Research Scientist
>Amgen Seattle
>-----Original Message-----
>From: histonet-bounces <@t> lists.utsouthwestern.edu
>[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Rebecca
>Sent: Thursday, December 07, 2006 1:34 PM
>To: histonet <@t> lists.utsouthwestern.edu
>Subject: [Histonet] GFP and B-Gal positive control slides
>Does anyone know where I can buy, or can suggest an easy way to make
>positive control slides for GFP and B-Gal.
>We have use AAV contstructs with either GFP or LAC-Z and injected
>into mice.  We have sectioned the tissue and would like to look for
>positive GFP or B-Gal.  It would be nice to have a positive control
>slide.  We plan on visualizing both with and without antibody
>amplification.  I would appreciate any suggestions.
>Thank You.
>Rebecca Nishi
>UC Irvine
>Christopher Reeve Foundation SCI Core/Anderson Lab
>1226 GNRF
>Irvine, CA 92697-4540
>rnishi <@t> uci.edu
>Phone/Fax: 949-824-9728
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>Histonet <@t> lists.utsouthwestern.edu
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Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367
406 994-4303 (FAX)

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