Immunoprecipitation of bovine brain membranes enriched in M1 and M2 muscarinic receptors with monoclonal antibody 10C7.
Monoclonal antibodies prepared against clathrin-coated vesicles cargo molecules were screened for their ability to precipitate the mAChR binding activity present in a light-density smooth membrane fraction from bovine brain called peak I. Only monoclonal antibody 10C7 was able to selectively sediment 38% (+/- 4.2) of the mAChR binding activity of peak I. Analysis by competition experiments of the supernatant obtained after the immunoprecipitation step reveals that neither the ratio of high/low affinity sites, nor the affinity ratio KH/KL was significantly altered for either of the muscarinic antagonists. The data implies that the bovine brain smooth-membrane compartment(s) expressing the mAb 10C7 antigen is also enriched in M1 and M2 mAChR.
Silva
W I
WI
Department of Pharmacology, Universidad Central del Caribe School of Medicine, Cayey, PR 00634.
Kohtz
D S
DS
Puszkin
S
S
D000818:Animals; D000911:Antibodies, Monoclonal; D001667:Binding, Competitive; D001921:Brain / Q000378:metabolism*; D002417:Cattle; D010890:Pirenzepine / Q000378:metabolism; D011233:Precipitin Tests; D011813:Quinuclidinyl Benzilate / Q000378:metabolism; D011976:Receptors, Muscarinic / Q000276:immunology* / Q000378:metabolism; D013347:Subcellular Fractions / Q000032:analysis
D000911:Antibodies, Monoclonal; D011976:Receptors, Muscarinic; D010890:Pirenzepine; D011813:Quinuclidinyl Benzilate