Detection of the M(r) 190,000 multidrug resistance protein, MRP, with monoclonal antibodies. 1994

D R Hipfner, and S D Gauldie, and R G Deeley, and S P Cole
Cancer Research Laboratories, Queen's University, Kingston, Ontario, Canada.

MRP is a M(r) 190,000 integral membrane phosphoglycoprotein that is overexpressed in some drug-selected resistant cell lines and has been shown to cause multidrug resistance in transfected cells. Five murine hybridoma cell lines (QCRL-1, QCRL-2, QCRL-3, QCRL-4, and QCRL-6) have been generated which secrete monoclonal antibodies (MAbs) that react specifically with membrane proteins of MRP-overexpressing, multidrug-resistant, drug-selected H69AR cells and MRP-transfected HeLa cells (T5) but not the respective parental (H69) and vector-transfected (C1) cells. The ability of three of these MAbs (QCRL-1, QCRL-2, and QCRL-3) to selectively immunoprecipitate a M(r) 190,000 protein from 35S-labeled H69AR and T5 membranes indicates that these MAbs are specific for MRP. MAb QCRL-1 is also capable of detecting the low levels of MRP present in revertant H69PR cells by immunoblot analysis. Indirect immunofluorescence analyses show that MAbs QCRL-1, QCRL-2, and QCRL-3) strongly and differentially react with fixed T5 and H69AR cells but not with unfixed cells, suggesting that these MAbs recognize intracellular MRP epitopes. The availability of reagents for the specific and sensitive immunodetection of MRP should greatly facilitate biological and clinical studies of this novel drug resistance protein.

UI MeSH Term Description Entries
D008856 Microscopy, Fluorescence Microscopy of specimens stained with fluorescent dye (usually fluorescein isothiocyanate) or of naturally fluorescent materials, which emit light when exposed to ultraviolet or blue light. Immunofluorescence microscopy utilizes antibodies that are labeled with fluorescent dye. Fluorescence Microscopy,Immunofluorescence Microscopy,Microscopy, Immunofluorescence,Fluorescence Microscopies,Immunofluorescence Microscopies,Microscopies, Fluorescence,Microscopies, Immunofluorescence
D008970 Molecular Weight The sum of the weight of all the atoms in a molecule. Molecular Weights,Weight, Molecular,Weights, Molecular
D006367 HeLa Cells The first continuously cultured human malignant CELL LINE, derived from the cervical carcinoma of Henrietta Lacks. These cells are used for, among other things, VIRUS CULTIVATION and PRECLINICAL DRUG EVALUATION assays. Cell, HeLa,Cells, HeLa,HeLa Cell
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D006825 Hybridomas Cells artificially created by fusion of activated lymphocytes with neoplastic cells. The resulting hybrid cells are cloned and produce pure MONOCLONAL ANTIBODIES or T-cell products, identical to those produced by the immunologically competent parent cell. Hybridoma
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D000911 Antibodies, Monoclonal Antibodies produced by a single clone of cells. Monoclonal Antibodies,Monoclonal Antibody,Antibody, Monoclonal
D000918 Antibody Specificity The property of antibodies which enables them to react with some ANTIGENIC DETERMINANTS and not with others. Specificity is dependent on chemical composition, physical forces, and molecular structure at the binding site. Antibody Specificities,Specificities, Antibody,Specificity, Antibody
D014407 Tumor Cells, Cultured Cells grown in vitro from neoplastic tissue. If they can be established as a TUMOR CELL LINE, they can be propagated in cell culture indefinitely. Cultured Tumor Cells,Neoplastic Cells, Cultured,Cultured Neoplastic Cells,Cell, Cultured Neoplastic,Cell, Cultured Tumor,Cells, Cultured Neoplastic,Cells, Cultured Tumor,Cultured Neoplastic Cell,Cultured Tumor Cell,Neoplastic Cell, Cultured,Tumor Cell, Cultured
D051379 Mice The common name for the genus Mus. Mice, House,Mus,Mus musculus,Mice, Laboratory,Mouse,Mouse, House,Mouse, Laboratory,Mouse, Swiss,Mus domesticus,Mus musculus domesticus,Swiss Mice,House Mice,House Mouse,Laboratory Mice,Laboratory Mouse,Mice, Swiss,Swiss Mouse,domesticus, Mus musculus

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