RE: amplification

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From:"Vaughan, Mary" <Mary.Vaughan@RoswellPark.org> (by way of histonet)
To:histonet@histosearch.com
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Good morning all,
	We did an amplified procedure a few years ago during the multi-drug
resistance craze....

Toth, K., Vaughan, M.M., Slocum, H.K.,et al. New Immunohistochemical
"Sandwich" Staining Method for MDR1 P-Glycoprotein Detection with JSB-1
Monoclonal Antibody in Formalin-Fixed, Paraffin-Embedded Human Tissues
Amer. J. Path. 144(2):227-236, 1994.

in brief ... used peroxidase labeled F[ab']2 fragment following the primary
AND following the PAP complex. The important thing with this assay was to
really have the primary concentration LOW and you must do an overnight [4C]
incubation. Also, used .03% Casein in all washes, and as the block.

I know, I know, it's a lot of steps..... but when all else had failed
everyone.... it worked BEAUTIFULLY. Check out the pictures in the original
publication.

Best Regards,
Mary Vaughan HT (ASCP)
Roswell Park Cancer Institute
Elm + Carlton Sts.   CDC-121
Buffalo, NY 14263


-----Original Message-----
From: Gayle Callis [mailto:uvsgc@msu.oscs.montana.edu]
Sent: Wednesday, November 10, 1999 3:29 PM
To: histonet@pathology.swmed.edu
Subject: amplification


found the technic using RxM IgG, then M xRat IgG interesting.  If you use
F(ab')2 fragments of the IgG, you may clean it up even more, to not bind
to Fc receptors on tissues.  We were thinking of trying this type of
technic also.

Gayle Callis




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