[Histonet] Anti-beta-galactosidase antibodies
mrl0627 <@t> mail.ecu.edu
mrl0627 <@t> mail.ecu.edu
Sat Nov 22 11:55:31 CST 2003
The easy answer is,I suppose, "because my thesis committee wants it that way" (smile). Working with hemizygote (lethal mutation in homozygotes) of transgenic mouse with LacZ insertion in front of proteoglycan of interest, enzyme histo equiv (maybe endogenous bgal plus weak signal in tissue) and have been doing immuno for core protein on Xgal reacted slides. Working fine. DAB looks beautiful next to Xgal blue. Complication is that the other grad students in our lab use flourescent antibodies and apparently are getting some kind of autoflourescent problem (don't know the details)when they use 40-1a on previously xgal reacted tissue. My advisor (who REALLY wants immuno and xgal on same sections) wants me to see how it 40-1a looks on my slides and if I still have the false positive problem. So trying to slay more than 1 dragon! Thanks for the reply. Maureen
-----Original Message-----
From: Sharon Cooperman <scoop <@t> mail.nih.gov>
To: mrl0627 <@t> mail.ecu.edu
Sent: Wed, 19 Nov 2003 09:40:27 -0500
Subject: Re: [Histonet] Anti-beta-galactosidase antibodies
No, sorry. Just out of curiousity, why do you want to do both the
enzymatic and IHC? Also, I think that if the enzyme isn't destroyed
by the reaction it might work - eg., I wouldn't expect it to work on
peroxidase, but I don't know if beta gal activity persists after the
reaction.
Sharon
>Sharon,
>Have you used this antibody on previously X-gal reacted tissue? We
>are using 40-1a. Thanks, Maureen Loomer, MS candidate at ECU
>
>-----Original Message-----
>From: Sharon Cooperman <scoop <@t> mail.nih.gov>
>To: t-kuzniar <@t> md.northwestern.edu
>Sent: Tue, 18 Nov 2003 18:34:04 -0500
>Subject: Re: [Histonet] Anti-beta-galactosidase antibodies
>
>Hi. Chemicon AB1211, ABC kit, HRP with DAB on formalin fixed
>paraffin embedded tissue. We used it on a transgenic mouse
>expressing Beta gal - pretty weak but detectable with ABC kit and
>HRP. I think most of the transgenics express weakly and I would
>consider using tyramide (TSA) if you need more signal because
>background is very low. I also think some of the Abcam anti-beta gal
>abs are pretty good (I don't know which ones, I borrowed them).
>
>Sharon
>
>>Has anyone had any experience with polyclonal antibody against
>>b-galactosidase? What reporter system have you used? Thank you.
>>Tom Kuzniar
>>
>>
>>
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>--
>Sharon Cooperman <scoop <@t> mail.nih.gov>
>NIH, NICHD, CBMB 301.435-7735
>Building 18T, room 101 301.402-0078 fax
>Bethesda, MD 20892
>
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--
Sharon Cooperman <scoop <@t> mail.nih.gov>
NIH, NICHD, CBMB 301.435-7735
Building 18T, room 101 301.402-0078 fax
Bethesda, MD 20892
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