Production of a specific antiserum to rat brain glutamic acid decar☐ylase by injection of an antigen-antibody complex

Content Provider	Semantic Scholar
Author	Oertel, Wolfgang Hermann Schmechel, Donald E. Tappaz, Marcel L. Kopin, Irwin J.
Copyright Year	1981
Abstract	Abstract Production of an antiserum specific to rat brain l -glutamic acid decar☐ylase is described, featuring the injection of an antigen-antibody complex. Partial purification of glutamate decar☐ylase was first achieved through differential centrifugation, ammonium sulfate fractionation, chromatography on Sephadex G-150, preparative isoelectric focusing in sucrose gradient and polyacrylamide gel electrophoresis for the production of a preliminary polyvalent, so called ‘trapping’ glutamate decar☐ylase antiserum in sheep. In rat brain homogenate supernatant this antiserum maximally inhibited glutamate decar☐ylase activity by 80% and totally precipitated enzyme activity on centrifugation. This antiserum, however, was polyvalent, as in crossed immunoelectrophoresis it detected four antigens in rat brain homogenate supernatant and three antigens in a partially purified preparation of glutamate decar☐ylase. One of these three precipitin lines could be radioactively labelled by 2-[ 3 H]-γ-acetylenic γ-aminobutyrate, an irreversible inhibitor of glutamate decar☐ylase. Injection of this antigen-antibody-precipitin line into a new non-immunized sheep yielded a new antiserum, which slightly inhibited but maximally precipitated 85% of the glutamate decar☐ylase activity in rat brain homogenate supernatant. In crossed immunoelectrophoresis the latter antiserum detected one antigen in the partially purified preparation of glutamate decar☐ylase. In crossed immunoelectrophoresis with intermediate gel the antiserum altered the mobility of a single antigen in brain homogenate supernatant. Thus, according to precipitating characteristics, the second antiserum appears to be specific to glutamate decar☐ylase.
Starting Page	2689
Ending Page	2700
Page Count	12
File Format	PDF HTM / HTML
DOI	10.1016/0306-4522(81)90113-5
PubMed reference number	7322358
Journal	Medline
Volume Number	6
Alternate Webpage(s)	https://api.elsevier.com/content/article/pii/0306452281901135
Alternate Webpage(s)	https://www.sciencedirect.com/science/article/pii/0306452281901135?dgcid=api_sd_search-api-endpoint
Alternate Webpage(s)	https://doi.org/10.1016/0306-4522%2881%2990113-5
Journal	Neuroscience
Language	English
Access Restriction	Open
Content Type	Text
Resource Type	Article