Rabbit polyclonal anti-Cap43 antibody preparation and preliminary purification of.doc

 PAGE \* MERGEFORMAT 3 Rabbit polyclonal anti-Cap43 antibody preparation and preliminary purification of [Abstract] Objective To prepare the rabbit polyclonal antibody anti-Cap43. Methods 30 amino acid peptide composed of semi-antigen and keyhole limpet hemocyanin (mcKLH) by carbodiimide (EDC) method chemical coupling into the full antigen (peptide-mcKLH) immune rabbit; Preparation of polyclonal rabbit anti-Cap4443 antibody; Dot-ELISA, detection of antibody titers, after bitter - ammonium sulfate method purified antibody, sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS) electrophoresis was detected by Coomassie brilliant blue, the purity of the purified . Results After 11 weeks of immunization are polyclonal rabbit anti-Cap43 body titer of 1:500. Conclusion EDC coupling chemistry and macromolecular hapten carrier protein mcKLH, making it both immunogenicity and immune response of complete antigen, used to immunize rabbits to obtain polyclonal antibodies. [Keywords:] semi-antigen Epidemiological and animal experiments have confirmed that nickel compounds can cause cancer, lung cancer, the main result. To further study the Cap43-specific cause lung cancer in nickel by the body’s tissue and serum expression levels and their characteristics, the present study, sulfide carbodiimide (EDC) to connect multiple peptide and macromolecular carrier protein way to make the composition of complete antigen, thus Preparation of anti-Cap43-free rabbit polyclonal antibodies for the study provide the basis of lung cancer caused by nickel. 1 Materials and methods 1  1 materials peptides synthesized by the Shanghai Health Public automatic instrument synthesis; EDC Kit (United States Pierce Company), complete and incomplete Freund’s adjuvant (Sigma, USA); HRP-goat anti-rabbit and DAB (Beijing Zhong Shan Golden Bridge Bio Technologies, Inc.); nitrate cellulose membrane (NC film), SephadexTMG-25 desalting column (Amersham Biosciences, Sweden), fetal calf serum,