【摘要】 MAP(multipleantigenicpeptide)多肽法具有快速、简便、成本低廉的特点.用MAP法,直接在多肽合成仪上合成Connexin31羧基端一个多肽片段(250-266AA)的MAP肽,经超滤器纯化后,免疫新西兰雄兔后采血检测、并纯化.经Westernblotting、细胞免疫荧光染色、免疫沉淀证实得到的抗体为抗间隙连接蛋白31的特异抗体.更多还原

【Abstract】 MAP peptide has been shown to produce strong immunogenic response.Sy nthesized the multiple antigenic peptide,then the purified multiple antigenic p eptide solution was mixed with Freund’s complete or incomplete adjuvant and immu nized rabbit to prepare specific polyclonal antibody against the topology of Con nexin31.A full-length human Connexin31 gene was cloned into expression vector,pcDNA3.1/myc-his(-)B,and transformed into HT1080 cell line stably or into He la cell transiently.Western blotting analysis and immunoprecipitation suggested that the polyclonal antibody recognize the fusion protein of the Connexin31-my c.Cell immunofluorescence staining shows the Connexin 31 localized at the site of the cell-cell contact gap junction.The antiserum against Connexin31 prepare d by multiple antigenic peptide has good specificity.更多还原