title: [Generation of mouse UBE2W antibody and analysis of UBE2W expression in mouse tissues].
abstract: Ubiquitin conjugating enzyme functions as the second enzyme required for protein ubiquitination and plays an important role in ubiquitin transferring and substrate specific recognition. UBE2W, a newly described member of E2 family, was formerly reported probably involving in phototransduction or retinal degeneration in Drosophila. In this study, we report that murine UBE2W harbors a typical UBC domain and is highly conserved in different vertebrate homologues. GST-tagged UBE2W was expressed in E. coli BL21 (DE3) and purified with GST affinity chromatography. Using this antigen, we generated and further separated rabbit polyclonal antibody of UBE2W, of which the activity and specificity were confirmed by immunoblotting of transiently expressed myc-UBE2W fusion protein. Wide expression of UBE2W was found in brain, muscle, heart, lung, liver, spleen, kidney and testis of mouse with the generated antibody, indicating the functional importance of this novel protein. Furthermore, the UBE2W highly expression was confined to the adult testis and was developmental stage-specific.
authors.affiliation: Beijing Institute of Biotechnology, Beijing 100071, China.
authors.forename: Yingying
authors.initials: Y
authors.lastname: Zhang
authors.forename: Hengqi
authors.initials: H
authors.lastname: Zhu
authors.forename: Lixia
authors.initials: L
authors.lastname: Zhao
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Zhou
authors.forename: Peitang
authors.initials: P
authors.lastname: Huang
mesh_terms: D000818:Animals; D000906:Antibodies / Q000378:metabolism; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D013737:Testis / Q000378:metabolism; D014018:Tissue Distribution; D044763:Ubiquitin-Conjugating Enzymes / Q000276:immunology* / Q000378:metabolism*
chemical_list: D000906:Antibodies; D000911:Antibodies, Monoclonal; D011993:Recombinant Fusion Proteins; D044763:Ubiquitin-Conjugating Enzymes