RE: [Histonet] GFP and B-Gal positive control slides
Not very if there is minimal target.
With that endothelium being so very, very thin where that particular
cassette expresses, you can see it, barely, without TSA. With TSA is
If wherever your b-gal is being expressed in your particular model, if it is
a nice big, plump cell you don't need amplification. But if your promoter
and particular experiment means there is just not a lot of b-gal there in
first place, need the TSA.
For your particular experiment, if robust target there, amplification not
needed. If cell type and or method of delivery makes target less robust,
the TSA works wonders.
From: Gayle Callis [mailto:email@example.com]
Sent: Friday, December 08, 2006 10:00 AM
To: Koelling, Ray; Histonet@lists.utsouthwestern.edu
Subject: RE: [Histonet] GFP and B-Gal positive control slides
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
>For the Tie-2/lac Z mice. Fix in formalin and process to paraffin as
>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.
>[mailto:firstname.lastname@example.org] On Behalf Of Rebecca
>Sent: Thursday, December 07, 2006 1:34 PM
>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.
>Christopher Reeve Foundation SCI Core/Anderson Lab
>Irvine, CA 92697-4540
>Histonet mailing list
>Histonet mailing list
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-4303 (FAX)
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