RE: cross-reactive Ab for NHP flow

From: Reed, Doug S Dr USAMRIID (Doug.Reed@DET.AMEDD.ARMY.MIL)
Date: Thu Mar 08 2001 - 14:16:34 EST


Just to throw my 2 cents in on the matter - we have employed the use of the
APC conjugates for anti-human CD8 and anti-human CD56 with APC for staining
with both cynomolgus and rhesus macaques. Both work just fine.

I would suggest that you look in Pharmingen's catalog at what antibodies
cross-react with nonhuman primates, then turn to the human antibody section
and pick the fluorophore you want. You will find that the "human" antibodies
are substantially cheaper than the "monkey" antibodies, even when they are
the same clone and the same fluorophore. So far the only antibody where that
does not apply is the CD3 clone which is not sold as a anti-human reagent.

-Doug Reed

-----Original Message-----
From: mandy cromwell [mailto:mandy_cromwell@hms.harvard.edu]
Sent: Wednesday, March 07, 2001 6:05 PM
To: cyto-inbox
Subject: cross-reactive Ab for NHP flow



Peter,

If you have a B-D Pharmingen catalog you may have noticed that none
of the Abs listed in the NHP section appear to be available as APC
conjugates. If you look up the same clones in the human section,
however, you will find that several clones are listed as APC
conjugates. For example, the CD2 clone RPA-2.10, CD8 clone RPA-T8,
CD20 clone 2H7, and CD69 clone FN50 all x-r with rhesus and are
listed in the human section conjugated to APC.

Also, the CD4 clone SK3 (rhesus x-r) has been listed in the (old) B-D
Biosciences immunocytometry  catalog as an APC conjugate (though for
some reason it isn't in the current B-D Pharmingen).

I don't know of any x-r CD28 or L-selectin clones that are available
in APC-maybe someone else can help out there.

Take a look at Keith Reiman's NHP reagent database for more
crossreactivity information:
http://rncc.bidmc.harvard.edu/labs/viral/Frameset_CD.html

Best regards,

Mandy
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
~~~
Mandy Cromwell, PhD
New England Regional Primate Research Center
Harvard Medical School
southborough, MA  01772
(508)624-8022


>I am currently setting up assays for measuring CD antigens on rhesus monkey
>T-cells by 4-color flow cytometry. I am trying to identify a source of
human
>Mabs (e.g. CD2, CD3, CD4, CD8, CD20, CD28, CD62L, CD69, etc) which are
>linked to APC and cross-react with rhesus T-cells.
>
>Your help would be greatly appreciated.
>
>Peter Silvera, PhD.
>Retrovirology Department
>Southern Research Institute
>431 Aviation Way
>Frederick, MD 21701
>Tel: 301-694-3232
>Fax: 301-694-7223
>e-mail: silvera@sri.org
>
>AND
>
>Henry Jackson Foundation
>HIV Research Program
>Rockville, MD 20850.

Mandy



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