Preparation and characterization of a novel monoclonal antibody specific to human CAD protein.
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.
Li
Mengbin
M
Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi'an, Shaanxi Province, China.
Zhang
Hongwei
H
Hong
Liu
L
Jin
Yan
Y
Gong
Taiqian
T
Sun
Li
L
Yang
Jianjun
J
Zheng
Xiushan
X
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
D000911:Antibodies, Monoclonal; C033626:CAD trifunctional enzyme; D007074:Immunoglobulin G; D001221:Aspartate Carbamoyltransferase; D004080:Dihydroorotase; D002223:Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing)