Characterization of monoclonal antibody to L-homocysteic acid and its immunohistochemistry in rat hippocampus.
L-homocysteic acid (HCA) and other amino acids were conjugated to rat brain material (extracted rat brain protein) with glutaraldehyde to form HCA- and amino acids-brain material conjugates. The specificity of monoclonal antibody (McAb) was tested on serial dilution test and absorption test on enzyme-linked immunosorbent assay (ELISA) using these conjugates as antigens instead of amino acids-BSA (bovine serum albumin) conjugates used previously. The characterized McAb was applied for immunohistochemical staining using PAP (peroxidase antiperoxidase) technique in combination with silver enhancement of diamino-benzene (DAB) products. The results indicated that McAb to L-HCA reacted with L-HCA-brain material conjugates, but not with other amino acids-brain material conjugates so far tested. McAb absorbed with L-HCA-brain material abolished or decreased immunoreactivity of L-HCA-brain material with McAb. The antibody selectively stained subpopulation of cells and processes in the hippocampus fixed with glutaradehyde. Absorption of McAb with L-HCA-brain material abolished immunohistochemical staining. These results suggested that McAb was specific for L-HCA-brain materials and could be used for imuno-histocytochemistry. This would provide a new tool for immunohistochemical visualization and localization of L-HCA in the nervous system.
Liu
Changjin
C
Department of Physiology, School of Basic Medical Sciences, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Li
Ai
A
Hu
Xinwu
X
Tang
Ming
M
Du
Yimei
Y
Zhang
Liangpin
L
Chen
Lei
L
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D006624:Hippocampus / Q000276:immunology / Q000378:metabolism*; D006710:Homocysteine / Q000031:analogs & derivatives* / Q000276:immunology; D007150:Immunohistochemistry; D051381:Rats; D017207:Rats, Sprague-Dawley
D000911:Antibodies, Monoclonal; D006710:Homocysteine; C007956:homocysteic acid