S-peptide epitope tagging for protein purification, expression monitoring, and localization in mammalian cells.
Epitope tags are widely used in cell biology and biochemistry research. The S-peptide/S-protein interaction has previously been utilized to purify polypeptides expressed in bacteria. We have now re-engineered the S-peptide/S-protein system to allow isolation of S-peptide-tagged polypeptides and their binding partners from eukaryotic cells with S-protein-agarose. In addition, two anti-S-peptide monoclonal antibodies have been generated for analysis of expression and subcellular localization of S-peptide-tagged polypeptides. These reagents make the S-peptide/S-protein system an attractive alternative to currently available epitope tagging methods.
Hackbarth
Jennifer S
JS
Mayo Clinic College of Medicine, Rochester, MN 55905, USA.
Lee
Sun-Hee
SH
Meng
Xue Wei
XW
Vroman
Benjamin T
BT
Kaufmann
Scott H
SH
Karnitz
Larry M
LM
D045744:Cell Line, Tumor; D018604:Epitope Mapping / Q000379:methods*; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D009363:Neoplasm Proteins / Q000302:isolation & purification* / Q000378:metabolism*; D010446:Peptide Fragments / Q000235:genetics* / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods; D025941:Protein Interaction Mapping / Q000379:methods*; D012259:Ribonuclease, Pancreatic / Q000235:genetics* / Q000378:metabolism*; D014018:Tissue Distribution
D009363:Neoplasm Proteins; D010446:Peptide Fragments; C060871:ribonuclease A S-peptide; D012259:Ribonuclease, Pancreatic