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Department of Pathology, Free University Hospital, De Boelelaan 1117, 1081 HV Amsterdam [M. J. F., M. A. I., G. L. S., J. M. F., C. J. L. M. M., R. J. S.], and Division of Molecular Biology, Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, [G. J. R. Z.] the Netherlands
We have generated rat and murine monoclonal antibodies against multidrug resistance-associated protein (MRP), a Mr 180,000–195,000 membrane glycoprotein involved in a non-P-glycoprotein multidrug resistance of human tumor cells. The antibodies were raised against two different segments of MRP and found to be suitable for protein blot analyses, immunohistochemical and cytochemical studies, as well as flow cytometry of permeabilized cells. The antibodies do not cross-react with the human P-glycoproteins. Immunocytochemistry using MRP-overexpressing tumor cells of different histogenetic origins showed that MRP is predominantly located in the plasma membrane. Immunoelectron microscopy confirmed the plasma membrane location of MRP. The MRP antibodies provide a sensitive and specific tool for studies on MRP-mediated multidrug resistance.
1 This work was supported by a Grant NKI 91-18 from the Dutch Cancer Society (to P. Borst and G. J. R. Z.). M. A. I. is a recipient of a European Society for Medical Oncology Fellowship.
2 To whom requests for reprints should be addressed.
Received 6/24/94. Accepted 7/21/94.
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M.L. den Boer, R. Pieters, K.M. Kazemier, M.M.A. Rottier, C.M. Zwaan, G.J.L. Kaspers, G. Janka-Schaub, G. Henze, U. Creutzig, R.J. Scheper, et al. Relationship Between Major Vault Protein/Lung Resistance Protein, Multidrug Resistance-Associated Protein, P-Glycoprotein Expression, and Drug Resistance in Childhood Leukemia Blood, March 15, 1998; 91(6): 2092 - 2098. [Abstract] [Full Text] [PDF]
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D. R. Hipfner, K. C. Almquist, E. M. Leslie, J. H. Gerlach, C. E. Grant, R. G. Deeley, and S. P. C. Cole Membrane Topology of the Multidrug Resistance Protein (MRP). A STUDY OF GLYCOSYLATION-SITE MUTANTS REVEALS AN EXTRACYTOSOLIC NH2 TERMINUS J. Biol. Chem., September 19, 1997; 272(38): 23623 - 23630. [Abstract] [Full Text] [PDF]
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M. Puder and R. J. Soberman Glutathione Conjugates Recognize the Rossmann Fold of Glyceraldehyde-3-phosphate Dehydrogenase J. Biol. Chem., April 18, 1997; 272(16): 10936 - 10940. [Abstract] [Full Text] [PDF]
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M. Gao, D. W. Loe, C. E. Grant, S. P.C. Cole, and R. G. Deeley Reconstitution of ATP-dependent Leukotriene C4 Transport by Co-expression of Both Half-molecules of Human Multidrug Resistance Protein in Insect Cells J. Biol. Chem., November 1, 1996; 271(44): 27782 - 27787. [Abstract] [Full Text] [PDF]
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Év. Bakos, T.ás Hegedüs, Z. Holló, E. Welker, Gáb. E. Tusnády, G. J. R. Zaman, M. J. Flens, A.ás Váradi, and B.áz. Sarkadi Membrane Topology and Glycosylation of the Human Multidrug Resistance-associated Protein J. Biol. Chem., May 24, 1996; 271(21): 12322 - 12326. [Abstract] [Full Text] [PDF]
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