RE: IHC with IgM (thanks!)

From:Marcia Bentz <mb7x@virginia.edu>

Thanks to all who responded to my question. I will call around and see if I
can find an anti-rat IgM secondary that is mouse adsorbed. Let you know how
it works out.

10:56 AM 10/19/2000 -0400, Connolly, Brett M wrote:
>Marcia,
>
>If it were me, I would go with the rat anti-human CD77 followed by a
>biotinylated anti-rat IgM that was preferably adsorbed against mouse tissue.
>Many companies offer anti-rat IgM (ie. Jackson ImmunoResearch, Biogenex,KPL,
>maybe Innogenex---Abizar are you there?). Call around and see if any are
>mouse adsorbed. I use an anti-rat IgG from Biogenex that is mouse adsorbed,
>maybe they have an IgM. 
>
>I don't think that rat IgG will bind the rat IgM, it depends on how purely
>it is prepared. However sometimes these secondaries are actually a mixture
>of IgA, IgM, and IgG). 
>
>I would also call tech services of the CD77 supplier and ask them if they
>have any data on crossreactivity with the antibody.
>
>Lastly, when you decide to enroll in another immunology class, sign your
>boss up too.
>
>Regards,
>Brett
>
>
>Brett M. Connolly, Ph.D.
>Merck Research Laboratories
>Department of Pharmacology
>WP26A-3000
>PO Box 4
>West Point, PA 19486
>Ph. 215-652-2501
>FAX 215-652-2075
>e-mail: brett_connolly@merck.com
>
>
>> ----------
>> From: 	Marcia Bentz[SMTP:mb7x@virginia.edu]
>> Sent: 	Thursday, October 19, 2000 6:48 AM
>> To: 	histonet@pathology.swmed.edu
>> Subject: 	IHC with IgM
>> 
>> Hello all,
>> Ended up dropping my immunology class several years ago due to a serious
>> illness and need the answer to a question. By boss is looking for an
>> antibody CD77 (if wishes could come true, anti-mouse). Two problems arise,
>> this is a IgM and I've only worked with IgG. I also only seem to be able
>> to
>> find mouse or rat anti-human CD77(I am staining mouse kidney). Since I use
>> the Vector Elite kits will the rat IgG work with an IgM primary?. My boss
>> insists it will, but well... he doesn't always know what he's talking
>> about
>> and has difficulty understanding IHC techniques sometimes.He also insists
>> there will be no cross reactions with mouse or rat proteins. Can anyone
>> answer this question for me?
>> 
>> Thanks!
>> 
>> Marcia
>> Marcia Bentz
>> Lab Specialist Sr
>> University of Virginia
>> Medicine/Nephrology
>> mb7x@virginia.edu
>> 804-982-1069
>> 
>> 
>
>
>
Marcia Bentz
Lab Specialist Sr
University of Virginia
Medicine/Nephrology
mb7x@virginia.edu
804-982-1069



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