A Rational Approach for Obtaining High-Specific Polyclonal Antibodies against Recombinant Alpha-Synuclein
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An approach for quick and efficient production of polyclonal antibodies to the target antigen, alpha-synuclein, has been proposed. Two methods have been employed to purify specific rabbit polyclonal antibodies against recombinant human alpha-synuclein, produced by subcutaneous immunization with complete Freund’s adjuvant. It was shown that purification on CNBr-activated Sepharose with immobilized alpha-synuclein resulted in antibody preparation with rabbit serum histidine-rich glycoprotein as a contaminant. Two-stage antibody purification procedure first on Sepharose with immobilized protein G, and then on alpha-synuclein immobilized column helps to avoid contamination and to obtain homogenous antibody preparation. Antibodies recognize different conformations of alpha-synuclein and can be used in a variety of immunochemical approaches, including immunocytochemistry.
Key words:polyclonal antibodies alpha-synuclein adjuvant immunization
- 1.Xiong, Y., Neifert, S., Karuppagounder, S.S., Stankowski, J.N., Lee, B.D., Grima, J.C., Chen, G., Ko, H.S., Lee, Y., Swing, D., Tessarollo, L., Dawson, T.M., and Dawson, V.L., Eneuro, 2017, vol. 4, no. 2, ENEURO.0004-17.2017. doi 10.1523/ ENEURO .0004-17.2017Google Scholar