Monoclonal antibody directed against glutaraldehyde conjugated glutamate and immunocytochemical applications in the rat brain.
Like other small-sized neurotransmitter molecules, glutamate (Glu) was conjugated to carrier proteins via glutaraldehyde (G). Human serum albumin (HSA) and thyroglobulin (TH) conjugates were alternately injected into mice. When a relevant immune response was obtained for antibody affinity and specificity, hybridization of spleen activated lymphocytes with SP2/O/Ag myeloma cells was performed. Supernatant culture media of hybridomas were tested for the presence of anti-conjugated Glu antibodies with our ELISA method. Selected hybridomas giving good antibody affinity and specificity were then cloned by the limiting dilution technique. Using DEAE-chromatographed ascites fluid, Glu reactivity was observed on the cortex and the hippocampus. Staining obtained with this monoclonal antibody was in agreement with that observed with previous polyclonal antisera directed against conjugated Glu or monoclonal anti-gamma-glutamyl-Glu antibody.
Chagnaud
J L
JL
Laboratoire d'immunologie, IBCN-CNRS, Université de Bordeaux II, France.
Campistron
G
G
Geffard
M
M
D000447:Aldehydes; D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis; D001921:Brain / Q000166:cytology / Q000378:metabolism*; D002540:Cerebral Cortex / Q000166:cytology / Q000378:metabolism; D004337:Drug Carriers; D005971:Glutamates / Q000276:immunology*; D018698:Glutamic Acid; D005976:Glutaral; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D051381:Rats; D011919:Rats, Inbred Strains; D012709:Serum Albumin; D013954:Thyroglobulin
D000447:Aldehydes; D000911:Antibodies, Monoclonal; D004337:Drug Carriers; D005971:Glutamates; D012709:Serum Albumin; D018698:Glutamic Acid; D013954:Thyroglobulin; D005976:Glutaral