Preparation and Application of Epitope Antibodies of Toxoplasma Actin
To prepare the antibodies against the epitopes of Toxoplasma actin(TgActin),the structure of TgActin were analyzed by BepiPred 1.0 Server and a specific polypeptide with a length of 12 amino acids was selected and synthesized for immunization of New Zealand rabbits.One week after the last immunization,serum samples were collected from the immunized rabbits and antibodies were purified by Protein G.The purity of antibodies was examined by SDS-PAGE and the titer was determined by ELISA.Moreover,the specificity of antibodies was analyzed by Western blot.Intracellular tachyzoites were detected by immunofluorescence using TgActin antibodies as the primary antibody.The results showed that the antibody titer was 1:128 000 in ELISA and the antibodies recognized TgActin in Western blotImmunofluorescence staining showed that the antibodies against TgActin bound to tachyzoites in parasitophorous vacuoles.In conclusion,the antibodies against the epitopes of TgActin were successfully prepared,and could be used as the internal reference antibody for the detection of T.gondii protein and immunofluorescence detection in cells.
Toxoplasma gondiiactinepitope antibodyloading control antibodyimmunofluorescence
国家科技期刊平台
NSTL
万方数据
