title: Tab2, a novel recombinant polypeptide tag offering sensitive and specific protein detection and reliable affinity purification.
abstract: The detection and purification of proteins are often time-consuming and frequently involve complicated protocols. The addition of a peptide tag to recombinant proteins can make this process more efficient. Many of the commonly used tags, such as Flagtrade mark, Myc, HA and V5 are recognized by specific monoclonal antibodies and therefore, allow immunoaffinity-based purification. Enhancing the current scope of flexibility in using diverse peptide tags, we report here the development of a novel, short polypeptide tag (Tab2) for detection and purification of recombinant proteins. The Tab2 epitope corresponds to the NH2-terminal seven amino acid residues of human TGFalpha. A monoclonal anti-Tab2 antibody was raised and characterized. To investigate the potential of this peptide sequence as a novel tag for recombinant proteins, we expressed several different recombinant proteins containing this tag in E. coli, baculovirus, and mammalian cells. The data presented demonstrates the Tab2 tag-anti-Tab2 antibody combination is a reliable tool enabling specific Western blot detection, FACS analysis, and immunoprecipitation as well as non-denaturing protein affinity purification.
authors.affiliation: Berlex Biosciences, Department of Systems Biology, Richmond, USA. kerstin.crusius@schering.de
authors.forename: Kerstin
authors.initials: K
authors.lastname: Crusius
authors.forename: Silke
authors.initials: S
authors.lastname: Finster
authors.forename: John
authors.initials: J
authors.lastname: McClary
authors.forename: Wei
authors.initials: W
authors.lastname: Xia
authors.forename: Brent
authors.initials: B
authors.lastname: Larsen
authors.forename: Douglas
authors.initials: D
authors.lastname: Schneider
authors.forename: Hong-Tao
authors.initials: HT
authors.lastname: Lu
authors.forename: Sara
authors.initials: S
authors.lastname: Biancalana
authors.forename: Jian-Ai
authors.initials: JA
authors.lastname: Xuan
authors.forename: Alicia
authors.initials: A
authors.lastname: Newton
authors.forename: Debbie
authors.initials: D
authors.lastname: Allen
authors.forename: Peter
authors.initials: P
authors.lastname: Bringmann
authors.forename: Ronald R
authors.initials: RR
authors.lastname: Cobb
mesh_terms: D020816:Amino Acid Motifs / Q000235:genetics; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology; D002478:Cells, Cultured; D002846:Chromatography, Affinity / Q000379:methods*; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D000939:Epitopes / Q000737:chemistry*; D005434:Flow Cytometry / Q000379:methods; D005822:Genetic Vectors / Q000138:chemical synthesis; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods; D007313:Insecta; D010446:Peptide Fragments / Q000235:genetics / Q000276:immunology*; D010770:Phosphotransferases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011485:Protein Binding / Q000276:immunology; D017434:Protein Structure, Tertiary / Q000235:genetics; D011956:Receptors, Cell Surface / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000138:chemical synthesis / Q000276:immunology / Q000302:isolation & purification*; D016211:Transforming Growth Factor alpha / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D010446:Peptide Fragments; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; D016211:Transforming Growth Factor alpha; D010770:Phosphotransferases