Immunoassay for the beta gamma subunits of GTP-binding proteins and their regional distribution in bovine brain.
Antibodies were raised in rabbits against the beta gamma subunits of bovine brain GTP-binding proteins, and were purified with a beta gamma-coupled Sepharose column. Purified antibodies reacted strongly with 36,000-dalton beta subunit and slightly with 35,000-dalton beta and gamma subunits, but not with other proteins in an immunoblot assay. Using these purified antibodies, a sensitive enzyme immunoassay method for the quantification of brain beta gamma was developed. The assay system consisted of polystyrene balls with immobilized antibody F(ab')2 fragments and the same antibody Fab' fragments labeled with beta-D-galactosidase from Escherichia coli. The minimum detection limit of the assay was 3 fmol, or 130 pg. Samples from various regions of bovine brain were solubilized with 2% sodium cholate and 1 M NaCl, and the concentrations of beta gamma were determined. The beta gamma were detected in all the regions, and the highest concentrations were observed in the cerebral cortex and nucleus caudatus. The concentrations of beta gamma were higher than those of alpha subunit of GTP-binding protein, Go, in all the regions.
D000818:Animals; D000906:Antibodies / Q000032:analysis; D001923:Brain Chemistry; D002417:Cattle; D002467:Cell Nucleus / Q000032:analysis; D002540:Cerebral Cortex / Q000032:analysis; D019204:GTP-Binding Proteins / Q000276:immunology / Q000302:isolation & purification*; D007124:Immunoenzyme Techniques; D011817:Rabbits
D000906:Antibodies; D019204:GTP-Binding Proteins