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Preparation and characterization of a novel monoclonal antibody specific to human CAD protein

Mengbin Li1, Hongwei Zhang, Liu Hong

  • 1Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi'an, Shaanxi Province, China.

Hybridoma (2005)
|March 3, 2009
PubMed

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.

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