Abstract
Russian 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. Predlozhena sovokupnost' priemov dlia bystrogo i prostogo polucheniia poliklonal'nykh antitel k tselevomu antigenu – al'fa-sinukleinu. Dlia ochistki spetsificheskikh poliklonal'nykh antitel krolika protiv rekombinantnogo al'fa-sinukleina cheloveka, narabotannykh v rezul'tate podkozhnoĭ immunizatsii s polnym ad"iuvantom Freĭnda, byli ispol'zovany dva metoda. Pokazano, chto ochistka na CNBr-aktivirovannoĭ sefaroze s immobilizovannym al'fa-sinukleinom privodit k polucheniiu preparata antitel, soderzhashchego v kachestve primesi obogashchennyĭ gistidinovymi ostatkami glikoprotein krovi krolika (histidine-rich glycoprotein). Dvukhstadiĭnaia ochistka spetsificheskikh antitel na sefaroze s immobilizovannym belkom G, a zatem na kolonke s immobilizovannym al'fa-sinukleinom pozvoliaet izbavit'sia ot primesnogo belka i poluchit' gomogennyĭ preparat antitel. Poluchennye antitela uznaiut razlichnye konformatsii al'fa-sinukleina i mogut byt' ispol'zovany dlia vsekh vidov immunnogo analiza, vkliuchaia immunotsitokhimicheskiĭ.