Immunochemical detection of the multidrug resistance-associated protein MRP in human multidrug-resistant tumor cells by monoclonal antibodies.
We have generated rat and murine monoclonal antibodies against multidrug resistance-associated protein (MRP), a M(r) 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.
Flens
M J
MJ
Department of Pathology, Free University Hospital, Amsterdam, The Netherlands.
Izquierdo
M A
MA
Scheffer
G L
GL
Fritz
J M
JM
Meijer
C J
CJ
Scheper
R J
RJ
Zaman
G J
GJ
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000818:Animals; D000911:Antibodies, Monoclonal; D002352:Carrier Proteins / Q000032:analysis; D004351:Drug Resistance; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007120:Immunochemistry; D008562:Membrane Glycoproteins / Q000032:analysis* / Q000737:chemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D009363:Neoplasm Proteins / Q000032:analysis* / Q000737:chemistry; D051381:Rats; D017208:Rats, Wistar; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000276:immunology; D014407:Tumor Cells, Cultured
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; D008562:Membrane Glycoproteins; D009363:Neoplasm Proteins; D011993:Recombinant Fusion Proteins