Help needed in MDR Study

From: Gary Warnes (Gary.warnes@cancer.org.uk)
Date: Thu Oct 10 2002 - 03:12:13 EST


Hi PS,
	I have used Rhodamine123 (FITC Channel) to measure P-gp
activity with MRK-16-PE mcab, seemed to work fine; we also used a
blocking P-gp ab in combination with Rhodamine123 that was reported
(see ref) . I think you can also use Ho33342 (UV Channel), because
the group I was working with also needed to use FITC & PE mcab too -
but we never got around to trying that combination.

Hope this is of some help.

Gary Warnes
FACSLAB
Cancer Research UK
London, UK

Heterozygous MDR3 missense mutation associated with intrahepatic
cholestasis of pregnancy: evidence for a defect in protein
trafficking.
Dixon PH, Weerasekera N, Linton KJ, Donaldson O, Chambers J, Egginton
E, Weaver J,Nelson-Piercy C, de Swiet M, Warnes G, Elias E, Higgins
CF, Johnston DG, McCarthy MI,Williamson C. Hum Mol Genet 2000 May
1;9(8):1209-17.

P.S.Manogaran wrote:-

We are studying Multi Drug Resistance (MDR) in our
leukemic   patients using P-gp antibody (17F9 clone
,Becton Dickinson). These patients cells , as many
reported, express low level of P-gp and perhaps is the
reason for  losing  some of the positive cases. Many
people are using MRK-16 and many other different
antibodies to measure P-gp. How different are these
antibodies to 17F9(BD)? We would like to know whether
anyone is using any other MDR related antibody alone
or in combination with P-gp to get a better picture of
MDR in Leukemic  patients. Is it possible to measure,
P-gp expression and   functional feature together in
the same tube,	using a functional dye which does not
overlap it's fluorescence with FITC and PE.

We are trying  to get MDR positives cells. We will
appreciate if  someone could provide us these cells or
  suggest us  a source from where I can get these cells
.We are also interested in finding    a  protocol  to
establish  these drug resistance cells , possibly from
  K562 cells.
--
Dr. Gary Warnes
FACS Laboratory
Cancer Research UK
44 Lincoln's Inn Fields
London	WC2A 3PX

P: 020 7269 3394
F: 020 7269 3100
mail to:gary.warnes@cancer.org.uk
Charity No: 1089464
OCTOBER IS BREAST CANCER AWARENESS MONTH
To find out more visit www.cancerresearchuk.org



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