Fw: [Histonet] CD79 alpha dilemma - ANSWER CORRECTION

[Jan Shivers]

Hello everyone - my apologies for posting a partially incorrect answer last week.  I was looking at the wrong line on a spreadsheet for CD79a abbreviated protocol information and wrote down the wrong HIER information.

What we do here currently with HIER for Biocare's CD79a (clone HM47/A9) on dogs, cats, pigs, cows, etc., is unmask antigens using:  0.1M Citrate buffer, pH 6.0 in a Biocare Decloaking Chamber.  (High pH retrieval solution does expose more antigenic sites, but it damages the tissue more when using the pressure cooker method.)  In the past we used high pH retrieval solution when doing HIER using a microwave oven (gentler method).

Jan Shivers
UMN Vet Diag Lab


----- Original Message ----- 
From: "Jan Shivers" <shive003 <@t> umn.edu>
To: "Farish, Craig" <CFarish <@t> csu.edu.au>; <histonet <@t> lists.utsouthwestern.edu>
Sent: Friday, May 13, 2011 9:14 AM
Subject: Re: [Histonet] CD79 alpha dilemma


> Hi Craig,
> 
> I have found CD79a from Biocare (cat. # CM067C) to stain dog, cat, pig, cow, 
> horse, sheep, and mouse so far.  Have not tried it on other species yet (in 
> other words, I don't have any negative species results to report).
> 
> I do have to use HIER with high pH retrieval buffer (Dako; contains EDTA) to 
> expose antigens (in a Biocare Decloaking Chamber/pressure cooker).  Citrate 
> buffer doesn't do the job well enough.  Microwave oven antigen retrieval 
> will also work, but it exposes fewer epitopes than the pressure cooker.
> 
> If you have any more questions, feel free to contact me directly.
> 
> Jan Shivers
> Senior Scientist
> IHC/Histology/EM Section Head
> Pathology Teaching Program
> University of Minnesota
> Veterinary Diagnostic Laboratory
> 1333 Gortner Ave.
> St. Paul, MN  55108
> 612-624-7297
> shive003 <@t> umn.edu
> 
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> 
> 
> ----- Original Message ----- 
> From: "Farish, Craig" <CFarish <@t> csu.edu.au>
> To: <histonet <@t> lists.utsouthwestern.edu>
> Sent: Thursday, May 12, 2011 8:35 PM
> Subject: [Histonet] CD79 alpha dilemma
> 
> 
> Hi folks - has anyone out there found a Cd79α clone and supplier which will 
> cover multiple species reliably?
> I can find clones which will stain cats and dogs (and humans), and clones 
> which will cover horses, pigs, cows and primates (and humans, but not 
> opossums for some reason??) but i'm yet to find one which will cross-react 
> with both the large and small animals commonly encountered in a vet lab. I 
> would rather not have to stock 2 versions of the same antibody if I can 
> avoid it.
> Any suggestions would be much appreciated.
> As always, thanks to everyone who contributes to histonet,
> Craig
> 
> Craig Farish
> Senior Technical Officer
> Veterinary Diagnostic Laboratory
> School of Animal and Veterinary Sciences
> Charles Sturt University
> Boorooma Street
> Wagga Wagga
> NSW 2678
> Australia
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