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

  • K. V. Barinova
  • A. K. Melnikova
  • E. V. Schmalhausen
  • V. I. Muronetz
Article
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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.

Key words:

polyclonal antibodies alpha-synuclein adjuvant immunization 

Notes

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Copyright information

© Pleiades Publishing, Ltd. 2018

Authors and Affiliations

  • K. V. Barinova
    • 1
    • 3
  • A. K. Melnikova
    • 2
  • E. V. Schmalhausen
    • 1
  • V. I. Muronetz
    • 1
    • 2
  1. 1.Belozersky Institute of Physico-Chemical Biology, Moscow State UniversityMoscowRussia
  2. 2.Faculty of Bioengineering and Bioinformatics, Moscow State UniversityMoscowRussia
  3. 3.Sechenov First Moscow State Medical University, Institute of Molecular MedicineMoscowRussia

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