Preparation of a polyclonal antibody against N-terminal of 1A6/DRIM with MAPs method / 北京大学学报(医学版)

ABSTRACT

Objective: To obtain the antibody against N terminal of 1A6/DRIM, and thereafter get the profile of 1A6/DRIM expression in different cell lines. Methods: The N terminal of 1A6/DRIM (aa 577 714) was cloned into pGEX 4T 3. Multiple antigenic peptides (MAPs)(aa638 661) was synthesized as the antigen with Fmoc/PyBOP method. Rabbits were immunized by injecting the MAPs and the immunized sera were analyzed with ELISA and Western Blot. The Western Blot and immunofluorescence were performed to analyze the expressing profiles of the 1A6/DRIM in different tumor cell lines. Results: The antibody specifically recognized the full length of 1A6/DRIM as a 310 kDa band, which was also recognized by C terminal monoclonal antibody shown by Western Blot. 1A6/DRIM is expressed in multiple tumor cell lines and mainly located in the nuclei. Conclusion: Preparation of the antibody with MAPs is a useful technique when the fusion proteins can not be induced in E coli . The antibody we got via MAPs has supplied a good tool for further studies on the functions of the novel gene 1A6/DRIM.