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. Author manuscript; available in PMC: 2013 Jul 1.
Published in final edited form as: Nat Chem Biol. 2012 Nov 11;9(1):43–50. doi: 10.1038/nchembio.1118

Figure 2. Characterization of 1 in cell lysate and live cells.

Figure 2

(a) HT-1 labels endogenous SFKs in cell lysates. Immunoblots with an anti-SFK antibody are shown for photo-crosslinking experiments performed with COS-7 and HeLa cell lysates. (b) Endogenous SFKs in live cells transiently expressing HaloTag are photo-crosslinked by 1. COS-7 cells transiently expressing HaloTag were treated with 1 and then irradiated with UV light. Immunoblotting with an anti-SFK antibody shows the presence of mass-shifted SFKs. Addition of an active site competitor prevents photo-crosslinking. (c) 1 labels nonreceptor tyrosine kinases in live cells. V5-tagged nonreceptor tyrosine kinases were coexpressed with HaloTag in COS-7 cells. After photo-crosslinking, the percent crosslinked kinase was determined with an anti-V5 antibody (mean ± SEM, n = 3).