Biotinylated Proteins Are Affinity Purified Using Immobilized Streptavidin and Analyzed (IMAGE)
Caption
(a) Streptavidin tetramer (left) was engineered to form a monomer that binds biotin with low nM affinity (right). (b) HEK293 cells displaying biotinylated CFP (top) were fluorescently labeled with Alexa 488 conjugated mSA (bottom). Fluorescent images taken at i) t = 0 and ii) 50 min indicate little dissociation of bound mSA. (c) mSA fused to the Z domain was used to form a noncovalent complex with antibody and recruit reactive biotin to a protein of interest, Erk-2. Biotinylated proteins are affinity purified using immobilized streptavidin and analyzed (e.g. SDS-PAGE, Western blot, mass spectrometry) to identify all proteins that interact with Erk-2.
Credit
TECHNOLOGY
Usage Restrictions
None
License
Licensed content