Generation of the antibody selectively against a novel thyroid hormone receptor beta isoform
A novel thyroid hormone receptor isoform TRβ△ was newly reported and deposited in Genbank(DQ191165.1) by our lab. To gain more understanding of this new variant. We raised rabbit polyclonal antibodies specifically against the characteristic epitope, amino acids 129-164 named as β△36, of TRβ△. Coding sequence for this β△36 peptide was obtained by PCR amplification and inserted into a GST fusion expression vector pGEX-3X. Recombinant plasmid was transformed into E. coli DH 5α and induced expression by 1 mmol/L IPTG at 25 degrees. Glutathione Sephorase 4B affinity chromatography purified fusion protein GST-β△ 36 was applied as immunogen to immune rabbits. An ELISA was developed to determine the titer of the antiserum. Rabbit anti GST-β△36 antiserum was collected when the titer reached 1:12000, and serum IgG was further purified by protein A affinity chromatography. Western blot was applied to testify the specificity of the antibody. The results showed that this rabbit anti-GST-β △ 36 polyclonal antibody could selectively recognize recombinant TRβ△ protein, but not cross react with recombinant TRβ1.
polyclonal antisurm thyroid hormone receptor isoform recombinan fusion protein
Ying Liu Bei Sun Yan Ma Liling Hu Xueqin Zhao Dongchun Liang Aijun Zuo Gang Guo Jingyu Zhang
Institute of Endocrinology and Metabolic Disease Hospital,Tianjin Medical University,Tianjin 300070, Institute of Endocrinology and Metabolic Disease Hospital,Tianjin Medical University Tianjin 300070,
国际会议
北京
英文
1-4
2009-06-11（万方平台首次上网日期，不代表论文的发表时间）