title: Preparation and characterization of a novel monoclonal antibody specific to human CAD protein.
abstract: We have efficiently generated the first monoclonal antibody against the cis-aconitic acid decarboxylase (CAD) protein, which is an enzyme of low stability. Hybridomas were screened by indirect enzyme-linked immunosorbent assay (ELISA) using either purified 6 x His-CAD fusion protein or purified 6 x His-ZNRD1 fusion protein as a control. One monoclonal antibody (MAb) named Z12 (IgG1), which is effective in detecting the recombinant and the cellular protein, was characterized by ELISA and Western immunoblotting. The CAD protein was also detected in gastric cancer and colon cancer tissues by immunohistochemical analysis. Thus, Z12 binds to native CAD protein and should be useful in studies of CAD protein function and expression.
authors.affiliation: Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi'an, Shaanxi Province, China.
authors.forename: Mengbin
authors.initials: M
authors.lastname: Li
authors.forename: Hongwei
authors.initials: H
authors.lastname: Zhang
authors.forename: Liu
authors.initials: L
authors.lastname: Hong
authors.forename: Yan
authors.initials: Y
authors.lastname: Jin
authors.forename: Taiqian
authors.initials: T
authors.lastname: Gong
authors.forename: Li
authors.initials: L
authors.lastname: Sun
authors.forename: Jianjun
authors.initials: J
authors.lastname: Yang
authors.forename: Xiushan
authors.initials: X
authors.lastname: Zheng
mesh_terms: D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D001221:Aspartate Carbamoyltransferase / Q000276:immunology*; D015153:Blotting, Western; D002223:Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing) / Q000276:immunology*; D045744:Cell Line, Tumor; D004080:Dihydroorotase / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D006825:Hybridomas / Q000276:immunology*; D007074:Immunoglobulin G / Q000276:immunology; D007150:Immunohistochemistry
chemical_list: D000911:Antibodies, Monoclonal; C033626:CAD trifunctional enzyme; D007074:Immunoglobulin G; D001221:Aspartate Carbamoyltransferase; D004080:Dihydroorotase; D002223:Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing)