[Polyclonal antibody preparation and expression pattern analysis of CARP]

The present study aims at preparation of CARP antibody and analysis of CARP expression pattern.Through bioinformatics analysis the antigenicity of CARP, 279 bp cDNA fragment coding CARP N-terminal 93 amino acid was amplified by RT-PCR from mouse heart RNA, then cloned into pET-28b to construct the prokaryotic expression pET 28b-CARP N93. The plasmid was transformed into E.coli(BL21). The target fusion protein was expressed with induction of IPTG, and purified by affinity chromatography. The antiserum against CARP was obtained from the mice immunized with CARP. CARP expression patterns were examined by Western blot and immunofluorescence.Prepared CARP antibody; CARP specific expression in mouse heart and mainly localized in the nucleus in rat cardiac primary cells.Successfully generated CARP antibody and examined the expression pattern. The present study has laid a foundation for functional study of CARP.