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  • Specific detection of multidrug resistance proteins MRP1, MRP2, MRP3, MRP5, and MDR3 P-glycoprotein with a panel of monoclonal antibodies.

Specific detection of multidrug resistance proteins MRP1, MRP2, MRP3, MRP5, and MDR3 P-glycoprotein with a panel of monoclonal antibodies.

Cancer research (2000-10-04)
G L Scheffer, M Kool, M Heijn, M de Haas, A C Pijnenborg, J Wijnholds, A van Helvoort, M C de Jong, J H Hooijberg, C A Mol, M van der Linden, J M de Vree, P van der Valk, R P Elferink, P Borst, R J Scheper
ABSTRACT

Tumor cells may display a multidrug resistance phenotype by overexpression of ATP binding cassette transporter genes such as multidrug resistance (MDR) 1 P-glycoprotein (P-gp) or the multidrug resistance protein 1 (MRP1). MDR3 P-gp is a close homologue of MDR1 P-gp, but its role in MDR is probably minor and remains to be established. The MRP1 protein belongs to a family of at least six members. Three of these, i.e., MRP1, MRP2, and MRP3, can transport MDR drugs and could be involved in MDR. The substrate specificity of the other family members remains to be defined. Specific monoclonal antibodies are required for wide-scale studies on the putative contribution of these closely related transporter proteins to MDR. In this report, we describe the extensive characterization of a panel of monoclonal antibodies (Mabs) detecting several MDR-related transporter proteins in both human and animal tissues. The panel consists of P3II-1 and P3II-26 for MDR3 P-gp; MRPr1, MRPm6, MRPm5, and MIB6 for MRP1; M2I-4, M2II-12, M2III-5 and M2III-6 for MRP2; M3II-9 and M3II-21 for MRP3; and M5I-1 and M5II-54 for MRP5. All Mabs in the panel appeared to be fully specific for their cognate antigens, both in Western blots and cytospin preparations, as revealed by lack of cross-reactivity with any of the other family members. Indeed, all Mabs were very effective in detecting their respective antigens in cytospins of transfected cell lines, whereas in flow cytometric and immunohistochemical analyses, distinct differences in reactivity and suitability were noted. These Mabs should become valuable tools in studying the physiological functions of these transporter proteins, in screening procedures for the absence of these proteins in hereditary metabolic (liver) diseases, and in studying the possible contributions of these molecules to MDR in cancer patients.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-MRP2 Antibody, clone M2 I-4, culture supernatant, clone M2 I-4, Chemicon®
Pricing and availability is not currently available.
Sigma-Aldrich
Anti-MDR3 Antibody, clone P3 II-26, culture supernatant, clone P3 II-26, Chemicon®
Pricing and availability is not currently available.
Sigma-Aldrich
Anti-MRP1 Antibody, clone MRPm5, clone MRPm5, Chemicon®, from mouse
Pricing and availability is not currently available.