Development of novel mouse hybridomas producing monoclonal antibodies specific to human and mouse nucleolar protein SURF-6.
SURF-6 is an evolutionarily conserved nucleolar protein that is important for cell viability; however, its function in mammals still remains uncertain. The aim of this study is to generate monoclonal antibodies to human SURF-6 protein suitable for fundamental and biomedical research. The full-size human SURF-6 was expressed as a recombinant GST-fusion protein and used as an antigen to generate monoclonal antibodies, S79 and S148, specific for SURF-6. The monoclonal antibody produced by hybridoma clone S79 specifically recognizes endogenous SURF-6 by Western and immunofluorescence analyses in various cultured human cells, and by immunohistochemistry in paraffin-embedded sections of human breast cancer samples. Moreover, S79 immunoprecipitates protein complexes containing SURF-6 from HeLa cells extracts. The antibody S79 recognizes SURF-6 only in human cells; however, the antibody produced by hybridoma clone S148 can detect SURF-6 of human and mouse origin. Monoclonal antibodies to the nucleolar protein SURF-6 described in this work can be a useful tool for studies of ribosome biogenesis in normal and cancer cells.
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 16/10 Miklukho-Maklaya, Moscow, Russia.
Mikhail A
MA
Polzikov
Maria Yu
MY
Kordyukova
Larisa E
LE
Zavalishina
Charalambos
C
Magoulas
Olga V
OV
Zatsepina
D016475:3T3 Cells; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D000918:Antibody Specificity; D000941:Antigens / Q000276:immunology; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000737:chemistry / Q000276:immunology; D005260:Female; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D051379:Mice; D009687:Nuclear Proteins / Q000276:immunology*; D016612:Paraffin Embedding; D011993:Recombinant Fusion Proteins / Q000276:immunology
D000911:Antibodies, Monoclonal; D000941:Antigens; D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; C484378:SURF6 protein, human; C098938:Surf6 protein, mouse