Keywords

Trim22, Peptide Antibody, Immunoaffinity Chromatography, Westernblot, Indirect Immunofluorescence

Abstract

Objective To prepare anti-TRIM22 polypeptide antibodies and identify their properties. Methods The polypeptide of 15 amino acids at the N terminus of TRIM22 (MDFSVKVDIEKEVTC) was coupled to keyhole limpet hemocyanin (KLH), and antiserum was prepared by immunizing New Zealand white rabbits; the TRIM22 polypeptide was coupled to Affi-Gel10 to prepare an immunoaffinity chromatography column. The rabbit antiserum was purified by affinity chromatography to obtain a TRIM22 polypeptide-specific antibody. ELISA was used to determine the antibody titer and Westernblot was used to identify the antibody specificity. The antibody was used to detect the intracellular localization of the TRIM22 protein through indirect immunofluorescence experiments. As a result, anti-TRIM22 specific antibodies were obtained. Westernblot results showed that the antibody could specifically recognize eukaryotic and prokaryotic expressed TRIM22 proteins. Indirect immunofluorescence experiments using this antibody found that TRIM22 protein was mainly located in the nucleus of HepG2 cells. Conclusion The preparation of this antibody provides a powerful tool for the functional study of TRIM22 molecules.

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Original research done by Gao Bo, Duan Zhijian, Gao Zhen, Hong Xiaowu, Xu Wei, Xiong Sidong

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