Strep/FLAG tandem affinity purification (SF-TAP) to study protein interactions.
In recent years, several methods have been developed to analyze protein-protein interactions under native conditions. One of them, tandem affinity purification (TAP), combines two affinity-purification steps to allow isolation of high-purity protein complexes. This unit presents a methodological workflow based on an SF-TAP tag comprising a doublet Strep-tag II and a FLAG moiety optimized for rapid as well as efficient tandem affinity purification of native proteins and protein complexes in higher eukaryotic cells. Depending on the stringency of purification conditions, SF-TAP allows both the isolation of a single tagged-fusion protein of interest and purification of protein complexes under native conditions.
Gloeckner
Christian Johannes
CJ
Helmholtz Zentrum München, Neuherberg, Germany.
Boldt
Karsten
K
Helmholtz Zentrum München, Neuherberg, Germany.|Technical University of Munich, Munich, Germany.
Ueffing
Marius
M
Helmholtz Zentrum München, Neuherberg, Germany.|Technical University of Munich, Munich, Germany.
D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods*; D003001:Cloning, Molecular; D015870:Gene Expression; D006801:Humans; D046912:Multiprotein Complexes / Q000737:chemistry / Q000302:isolation & purification*; D009842:Oligopeptides; D010455:Peptides; D011485:Protein Binding; D011993:Recombinant Fusion Proteins; D014162:Transfection
C099517:Ala-Trp-Arg-His-Pro-Gln-Phe-Gly-Gly; D046912:Multiprotein Complexes; D009842:Oligopeptides; D010455:Peptides; D011993:Recombinant Fusion Proteins; C085706:FLAG peptide
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