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Streptavidin Bead Pulldown Assay to Determine Protein Homooligomerization.

Bio-protocol.. 2017-11; 
Ting-Hai Xu, Yan Yan, Kaleeckal G. Harikumar, Laurence J. Miller,Karsten Melcher,H. Eric Xu.
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Protein and Antibody Isolation … 15. Streptavidin MagBeads (GenScript, catalog number: L00424) 16 … proteins and associated C99(WT)-TEV site-rTA-Flag were recovered on Streptavidin MagBeads (GenScript) and eluted with SDS sample buffer, while the dimer interface mutants (T43P/V45P … Get A Quote

Abstract

Pulldown assay is a conventional method to determine protein-protein interactions in vitro. Expressing a protein of interest with two different tags allows testing whether both versions can be captured via one of the two tags as homooligomeric complex. This protocol is based on streptavidin bead capture of a biotinylated protein and co-associated Flag-tagged protein using Streptavidin MagBeads.

Keywords

Pulldown assay, Protein-protein interactions, Streptavidin bead, Biotinylated protein, Homooligomeric complex
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