Preparation and Characterization of Monoclonal Antibody against Human Mitochondrial Ribosomal Protein S17

Abstract: BALB/c mice were immunized with purified mitochondrial ribosomal protein S17(MRPS17).Spleen cells were fused with SP2/0 cells using polyethylene glycol and hybridoma cells were selected by HAT medium and screened with ELISA. After rounds of cloning with limited dilution method， one hybridoma cell line was obtained which stably secret monoclonal antibody of IgG2a. The obtained antibody was used for samples assay by Western blot， immunohistochemical and immunofluorescence staining. Western blot showed that the antibody specifically recognized about 13 kDa protein. Immunohistochemical staining showed strong positive signals located in the cytoplasm of muscle cells and melanoma cells. Immunofluorescence staining showed green fluorescence mainly in the formation of a spotted distribution inside the cytoplasm of HeLa cells， which was identical to the distribution of mitochondria shown by mitochondrion-selective stains (MitoTracker Redcm-H2XRos). These results indicated that a MRPS17 monoclonal antibody with high specificity was developed， which could be used in Western blot， immunohistochemical and immunofluorescent techniques.
    

CSTR: 32200.14.cjcb.2006.02.0019