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. 2009 Apr 24;4(4):e5315. doi: 10.1371/journal.pone.0005315

Figure 1. Purified SVOP-FLAG fusion proteins are labeled with 8-azido-ATP-biotin.

Figure 1

Recombinant SVOP-FLAG fusion protein was purified from transfected HEK293 cells with Anti-FLAG M2 affinity gel. About 5 µg protein preparation was used in each photoaffinity labeling reaction with 100 µM 8-azido-ATP-biotin in the presence or absence of 1 mM non-photoreactive ATP. A control without UV photolysis was set up in parallel. The samples were resolved by SDS-PAGE and transferred to PVDF membrane for western blot analysis. The bound 8-azido-ATP was visualized by ExtrAvidin-HRP and anti-FLAG antibody was used to detect the proteins.