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目的:制备兔抗人转录因子TFDP3、TFDP1的多克隆抗体,并鉴定其反应性及特异性。方法:选择TFDP3和TFDP1差异较大肽段,利用人工合成多肽免疫家兔,采用真核表达载体瞬时转染HEK293细胞表达TFDP3和TFDP1真核蛋白,通过Western blot分析抗体的反应性和特异性。结果:通过氨基酸序列比对选择TFDP3 17-26,TFDP1 17-32氨基酸残基合成多肽,制备了兔抗人TFDP3、TFDP1多肽抗体,Western blot检测显示,抗TFDP3抗体可以特异性识别TFDP3,而抗TFDP1抗体既可识别TFDP1又可与TFDP3结合。结论:制备了兔抗人TFD31和TFDP1的多肽抗体,且TFDP3抗体与TFDP1无交叉反应。
Objective: To prepare polyclonal antibodies against TFDP3 and TFDP1 and to identify their reactivity and specificity. Methods: The larger peptides of TFDP3 and TFDP1 were selected. Rabbit was immunized with synthetic peptides. The eukaryotic expression vectors of TFDP3 and TFDP1 were transiently transfected into HEK293 cells by eukaryotic expression vector. The reactivity and specificity of the antibodies were analyzed by Western blot. Results: The polypeptides of TFDP3 17-26 and TFDP1 17-32 amino acid residues were synthesized by amino acid sequence alignment. Anti-human TFDP3 and TFDP1 peptide antibodies were prepared by Western blot. Western blot showed that anti-TFDP3 antibody could specifically recognize TFDP3, The TFDP1 antibody recognizes both TFDP1 and TFDP3. Conclusion: Rabbit anti-human TFD31 and TFDP1 polypeptide antibodies were prepared, and TFDP3 antibody did not cross-react with TFDP1.