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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