HIGHLY SENSITIVE IMMUNOASSAY FOR THE ALPHA-SUBUNIT OF THE GTP-BINDING PROTEIN GO AND ITS REGIONAL DISTRIBUTION IN BOVINE BRAIN
HIGHLY SENSITIVE IMMUNOASSAY FOR THE ALPHA-SUBUNIT OF THE GTP-BINDING PROTEIN GO AND ITS REGIONAL DISTRIBUTION IN BOVINE BRAIN
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DOI：
10.1111/j.1471-4159.1987.tb05710.x
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发表时间：
1987-05-01
影响因子：
4.7
通讯作者：
KATO, K
中科院分区：
文献类型：
--
作者：
ASANO, T;SEMBA, R;KATO, K
Antisera were raised in rabbits against the .alpha. subunit of a GTP-binding protein, Go. Because the antisera cross-reacted weakly with the .alpha. subunit of inhibitory GTP-binding protein of adenylate cyclase (Gi), they were purified with a Go.alpha.-coupled Sepharose column. Purified antibodies reacted only with Go.alpha. and did not a cross-react with the Gi.alpha. subunit or .beta..gamma. subunits in an immunoblot assay. Using these purified antibodies, a highly sensitive enzyme immunoassay method for the quantification of bovine brain Go.alpha. 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 minimal detection limit of the assay was 0.1 fmol, or 4 pg. The assay was specific for Go.alpha., and it did not cross-react with Gi.alpha. or .beta..gamma.. Samples from various regions of bovine brain were solubilzed with 2% sodium cholate and 1 M NaCl, and the concentrations of Go.alpha. were determined. Go.alpha. was detected in all the regions, and the highest concentration was observed in the cerebral cortex. The immunohistochemical study showed that the neuropil was rich in Go.alpha.
