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[A rational approach to obtaining high-specific polyclonal antibodies against recombinant alpha-synuclein]

K V Barinova1, A K Melnikova2, E V Schmalhausen3

  • 1Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, Russia; Sechenov First Moscow State Medical University, Institute of Molecular Medicine, Moscow, Russia.

Biomeditsinskaia Khimiia
|July 3, 2018
PubMed

The approach for the quick and efficient production ofpolyclonal antibodies tothe 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.

Keywords:
adjuvantalpha-synucleinimmunizationpolyclonal antibodies

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