Characterization of Monoclonal Antibody to L-Homocysteic Acid and Its Immunohistochemistry in Rat Hi

来源 :华中科技大学学报(医学英德文版) | 被引量 : 0次 | 上传用户:apworld
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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 con-jugates. The specificity of monoclonal antibody (McAb) was tested on serial dilution test and ab-sorption 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) tech-nique in combination with silver enhancement of diamino-benzene (DAB) products. The results in-dicated that McAb to L-HCA reacted with L-HCA-brain material conjugates, but not with other a-mino acids-brain material conjugates so far tested. McAb absorbed with L-HCA-brain material a-bolished or decreased immunoreactivity of L-HCA-brain material with McAb. The antibody selec-tively 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 locali-zation of L-HCA in the nervous system.
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