title: Domain-Specific Monoclonal Antibodies Against Human Rev-erbβ.
abstract: The nuclear receptor Rev-erbβ is a potent transcriptional factor whose functional study has been limited by the lack of suitable antibodies against it. To better understand Rev-erbβ's biological roles, we generated five hybridoma cell lines secreting antibodies against human Rev-erbβ in mice immunized with the purified, prokaryotically expressed recombinant Rev-erbβ-6His fusion protein. Using Western blotting and immunofluorescence analyses, all the five monoclonal antibodies (MAbs) showed strong immunoreactivity to both prokaryotically and eukaryotically expressed recombinant Rev-erbβ. An immunoprecipitation study showed that all five monoclonal antibodies against Rev-erbβ were able to pull down the recombinant Rev-erbβ-Flag protein, but only one of the MAbs against Rev-erbβ, 37H8, could pull down the endogenous Rev-erbβ protein. Furthermore, domain specificity of these MAbs was characterized. Due to the high similarities between Rev-erbα and Rev-erbβ in the C and E domains, those C and E domain-specific anti-Rev-erbβ antibodies can react with human Rev-erbα as well. The MAbs produced in the study will provide a valuable tool for investigating the function of Rev-erbβ.
authors.affiliation: Laboratory of Gene Therapy, Department of Biochemistry, College of Life Sciences, Shaanxi Normal University, 199 South Chang'an Road, Xi'an, 710062, Shaanxi, People's Republic of China.|School of Modern Agriculture and Biotechnology, Ankang University, Ankang, 725000, Shaanxi, People's Republic of China.
authors.forename: Fang
authors.initials: F
authors.lastname: Chen
authors.affiliation: Laboratory of Gene Therapy, Department of Biochemistry, College of Life Sciences, Shaanxi Normal University, 199 South Chang'an Road, Xi'an, 710062, Shaanxi, People's Republic of China.
authors.forename: Yanqing
authors.initials: Y
authors.lastname: Li
authors.affiliation: Laboratory of Gene Therapy, Department of Biochemistry, College of Life Sciences, Shaanxi Normal University, 199 South Chang'an Road, Xi'an, 710062, Shaanxi, People's Republic of China.
authors.forename: Junli
authors.initials: J
authors.lastname: Zhao
authors.affiliation: Department of Pathology, Northwestern University Feinberg School of Medicine, Chicago, IL, 60611, USA.
authors.forename: Qinwen
authors.initials: Q
authors.lastname: Mao
authors.affiliation: Laboratory of Gene Therapy, Department of Biochemistry, College of Life Sciences, Shaanxi Normal University, 199 South Chang'an Road, Xi'an, 710062, Shaanxi, People's Republic of China. hbxia2001@163.com.
authors.forename: Haibin
authors.initials: H
authors.lastname: Xia
mesh_terms: D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000737:chemistry / Q000276:immunology*; D000918:Antibody Specificity; D057809:HEK293 Cells; D056945:Hep G2 Cells; D006801:Humans; D051379:Mice; D000072417:Protein Domains; D018160:Receptors, Cytoplasmic and Nuclear / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D012097:Repressor Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; C090977:NR1D2 protein, human; D018160:Receptors, Cytoplasmic and Nuclear; D012097:Repressor Proteins
keywords: Domain; Hybridoma; Monoclonal antibody; Rev-erbα; Rev-erbβ