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

   


1. Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, Russia; Sechenov First Moscow State Medical University, Institute of Molecular Medicine, Moscow, Russia
2. Faculty of Bioengineering and Bioinformatics, Lomonosov Moscow State University, Moscow, Russia
3. Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, Russia
4. Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, Russia; Faculty of Bioengineering and Bioinformatics, Lomonosov Moscow State University, Moscow, Russia
Type: Experimental study
DOI: 10.18097/PBMC20186403276      PubMed Id: 29964265
Year: 2018 vol: 64  issue:3  pages: 276-282
Abstract: 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.
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Reference: Barinova K.V., Melnikova A.K., Schmalhausen E.V., Muronetz V.I., A rational approach to obtaining high-specific polyclonal antibodies against recombinant alpha-synuclein, Biomeditsinskaya khimiya, 2018, vol: 64(3), 276-282.
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