Mouse anti-human cytochrome P450 1A2 synthetic peptide antiserum and identification of.doc

 PAGE \* MERGEFORMAT 11 Mouse anti-human cytochrome P450 1A2 synthetic peptide antiserum and identification of Author: Wu Zheng-Hua, Xiao-Bo Tang Wang Chih-kang Zhu Ling Ji Hongyu Zhao Camp [Keywords:] CYP1A2;, synthetic peptide;, antibody;, mouse [Abstract] Objective: To prepare mouse anti-human cytochrome P450 1A2 (CYP1A2) antibody and its characterization. Methods: Using bioinformatics analysis of CYP1A2 protein sequence, according to four indicators (hydrophilic, flexibility, antigenicity and surface nature) Select For synthetic peptide sequence, design and synthesis of CYP1A2 peptides. Them with carrier protein keyhole limpet hemocyanin (keyhole limpet hemocyanin, KLH) conjugate, the immune BALB / c female mice were prepared by anti-CYP1A2 antibodies. Indirect ELISA, titer, Western blot identification of their specificity. Results: The success of access for small molecules CYP1A2 antibodies. The antibody titer of 1:16000, with CYP1A2 synthetic peptide and natural CYP1A2 specific reaction occurs. Western blot results showed that the antibodies recognized the corresponding antigen relative molecular mass (Mr) of 58,000. Conclusion: The synthetic peptide KLH complex is immunogenic, can be used to prepare the appropriate antibodies. Preparation of synthetic peptide of mouse anti-CYP1A2 antibody titer and specificity of good. [Keywords:] CYP1A2; synthetic peptide; antibody; mice CYP1A2 and CYP1A1 is the CYP1A subfamily in the only two members of the [1], they are the expression in the human body has its own tissue specificity, CYP1A2 expression primarily in the liver [2], are also detected in brain tissues and CYP1A2 mRNA no reports of protein expression [3]. Due to CYP1A2 certain metabolic activation of heterocyclic aromatic carcinogens into carcinogens before, such as the CYP1A2 catalyzed the process of cooking the food produced in 2-Amino-1-methyl-6-phenyl-imino [4, 5b] pyridine hydroxylation of N2 activation reaction, in the chemical carcinogens plays