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A Rational Approach for Obtaining High-Specific Polyclonal Antibodies against Recombinant Alpha-Synuclein

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Abstract

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.

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Authors and Affiliations

  1. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Leninskie Gory 1 bldg 40, 119234, Moscow, Russia

    K. V. Barinova, E. V. Schmalhausen & V. I. Muronetz

  2. Faculty of Bioengineering and Bioinformatics, Moscow State University, Moscow, Russia

    A. K. Melnikova & V. I. Muronetz

  3. Sechenov First Moscow State Medical University, Institute of Molecular Medicine, Moscow, Russia

    K. V. Barinova

Authors
  1. K. V. Barinova
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  2. A. K. Melnikova
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  3. E. V. Schmalhausen
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  4. V. I. Muronetz
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Correspondence to V. I. Muronetz.

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Translated by A. Medvedev

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Barinova, K.V., Melnikova, A.K., Schmalhausen, E.V. et al. A Rational Approach for Obtaining High-Specific Polyclonal Antibodies against Recombinant Alpha-Synuclein. Biochem. Moscow Suppl. Ser. B 12, 316–321 (2018). https://doi.org/10.1134/S1990750818040029

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  • DOI: https://doi.org/10.1134/S1990750818040029

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