Immunoprecipitation of bovine brain membranes enriched in M1 and M2 muscarinic receptors with monoclonal antibody 10C7

W. I. Silva; D. S. Kohtz; S. Puszkin

doi:10.1016/0304-3940(90)90500-9

Immunoprecipitation of bovine brain membranes enriched in M1 and M2 muscarinic receptors with monoclonal antibody 10C7

W. I. Silva, D. S. Kohtz, S. Puszkin

Research output: Contribution to journal › Article › peer-review

Abstract

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 K_H K_L 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.

Original language	English
Pages (from-to)	89-94
Number of pages	6
Journal	Neuroscience Letters
Volume	113
Issue number	1
DOIs	https://doi.org/10.1016/0304-3940(90)90500-9
State	Published - May 18 1990

Keywords

Clathrin-coated vesicles
Immunoprecipitation assays
Membrane compartments
Monoclonal antibodies
Muscarinic receptors
Subcellular pathways

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10.1016/0304-3940(90)90500-9

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title = "Immunoprecipitation of bovine brain membranes enriched in M1 and M2 muscarinic receptors with monoclonal antibody 10C7",

abstract = "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.",

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T1 - Immunoprecipitation of bovine brain membranes enriched in M1 and M2 muscarinic receptors with monoclonal antibody 10C7

AU - Silva, W. I.

AU - Kohtz, D. S.

AU - Puszkin, S.

PY - 1990/5/18

Y1 - 1990/5/18

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AB - 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.

KW - Clathrin-coated vesicles

KW - Immunoprecipitation assays

KW - Membrane compartments

KW - Monoclonal antibodies

KW - Muscarinic receptors

KW - Subcellular pathways

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Immunoprecipitation of bovine brain membranes enriched in M1 and M2 muscarinic receptors with monoclonal antibody 10C7

Abstract

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