[Histonet] Anti-beta-galactosidase antibodies

Gayle Callis gcallis <@t> montana.edu
Wed Nov 19 09:59:20 CST 2003


Good point and my thoughts exactly.  I would certainly try the BETA GAL
Roche kit to develop lovely beta gal blue color before going to the
antibody.  I think one key thing is to have a good positive tissue control.
 Robin Taylor from Genentech sent me a wonderful protocol using this kit, I
am sure she would not mind if I shared it privately. It is very straight
forward.  The only time we used the antibody was to detect beta gal for
immunofluorescence.  There ARE fluorescent beta gal substrates available
but using antibody staining seems just as easy for this application. 

At 09:40 AM 11/19/2003 -0500, you wrote:
>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 &lt;scoop <@t> mail.nih.gov&gt;
>>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
>>
>>&gt;Has anyone had any experience with polyclonal antibody against
>>&gt;b-galactosidase? What reporter system have you used? Thank you.
>>&gt;Tom Kuzniar
>>&gt;
>>&gt;
>>&gt;
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>>
>>--
>>Sharon Cooperman      	     &lt;scoop <@t> mail.nih.gov&gt;
>>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
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>




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