Tab2, a novel recombinant polypeptide tag offering sensitive and specific protein detection and reliable affinity purification.
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.
Crusius
Kerstin
K
Berlex Biosciences, Department of Systems Biology, Richmond, USA. kerstin.crusius@schering.de
Finster
Silke
S
McClary
John
J
Xia
Wei
W
Larsen
Brent
B
Schneider
Douglas
D
Lu
Hong-Tao
HT
Biancalana
Sara
S
Xuan
Jian-Ai
JA
Newton
Alicia
A
Allen
Debbie
D
Bringmann
Peter
P
Cobb
Ronald R
RR
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
D000911:Antibodies, Monoclonal; D000939:Epitopes; D010446:Peptide Fragments; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; D016211:Transforming Growth Factor alpha; D010770:Phosphotransferases