RE: multi drug resistance protein

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From:"Vaughan, Mary" <Mary.Vaughan@RoswellPark.org>
To:'David Grehan' <david.grehan@olhsc.ie>, histonet@pathology.swmed.edu
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Good Morning David,
	I've done several years of Multi Drug Resistance Protein work. It
was a tough nut to crack back a few years ago. We use JSB-1 primary antibody
[far superior to C219] and the procedure is long... we've added extra IgG
fragments to give the DAB some extra places to bind. It's really a nice,
clean, amplified method. I can send you the worksheet for the procedure if
you'd like. Please find the related publications below (methods paper is
#2). I think #3 may be of special interest to you.

1)Toth, K., Vaughan, M.M., Slocum, H.K., Fredericks, W.J., Chen, Y.,
Arredondo, M.A., Harstrick, A., Karakousis, C., Baker, R.M., Rustum, Y.M.
Comparison of an Immunoperoxidase "sandwich" Staining Method and Western
Blot Detection of P-glycoprotein in Human Cell Lines and Sarcomas. Amer. J.
Path. 140:1009-1016, 1992.

2)Toth, K., Vaughan, M.M., Slocum, H.K., Arredondo, M.A., Takita, H., Baker,
R.M., Tsuro, T., and Rustum, Y.M. 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.

3)Toth, K., Vaughan, M.M., Peress, N.S., Slocum, H.K., Rustum, Y.M.  MDR1
p-glycoprotein is expressed by endothelial cells of newly-formed capillaries
in human gliomas but is not expressed in the neovasculature of other primary
tumors. Amer J. Path. 149(3): 853-858, 1996.

4)Toth, K., Vaughan, M.M., Schwartz, G., Winston, J.S., Skenderis, B.S.,
Slocum, H.K., Rustum, Y.M., Expression of the MRP and MDR1 Multidrug
Resistance Gene Products in 160 Untreated Human Carcinomas Studied by
Immunohistochemical Methods in Formalin-Paraffin Sections:.Int. J Surgical
Pathology 6(3):145-154, 1998

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

-----Original Message-----
From: David Grehan [mailto:david.grehan@olhsc.ie]
Hi histolanders,
I am trying to locate a good antibody to Multi Drug Resistance Protein
that works well on paraffin processed neuroblastoma tumours. Does anyone
have any experience using these antibodies with this tumour or any other
tumour type. I have tried a monoclonal antibody to MRP (QCRL1) from ID
Labs at a 1:20 dilution following pressure cooker AR without much
success. The MRP gene is over expressed in MYCN amplified  neuroblastoma
tumours .
Secondly does anyone routinely detect the amplified MYCN gene copy in
paraffin sections and where do you obtain the probe.

thanks.





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