Lacava J, et al. (2013) Improved native isolation of endogenous Protein A-tagged protein complexes. Biotechniques 54(4):213-6
Abstract: Here we report a modified peptide reagent useful for the rapid, native elution of protein complexes containing a Protein-A-tagged component. We tested this reagent for the elution of tagged endogenous protein complexes from yeast (Nup53p/Nup170p dimer; Nup1p/Kap95p/Kap60p trimer; pentameric GINS complex) and bacteria (RNAPII holoenzyme). The majority of the affinity-isolated material is released within 15 minutes under mild conditions, and the elution reagent itself is readily depleted from the elution mixture by simple spin column gel filtration. This reagent is ideal for eluting protein complexes after Protein A / IgG affinity isolation when protease cleavage is not possible or not desirable and facile depletion of the elution reagent is needed.
|Status: Published||Type: Journal Article | Research Support, N.I.H., Extramural||PubMed ID: 23581468|
Topics addressed in this paper
Number of different genes curated to this paper: 5
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