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. Author manuscript; available in PMC: 2011 Mar 28.
Published in final edited form as: Nat Chem Biol. 2010 Dec 12;7(1):58–63. doi: 10.1038/nchembio.495

Figure 3. Verification of lysine succinylation by western blot analysis.

Figure 3

(a) Specificity of anti-SuccK antibody using dot-spot assays. Peptide libraries bearing a fixed unmodified lysine, acetyllysine or succinyllysine were spotted on nitrocellulose membrane with 10-fold dilutions. The 13-residue randomized peptide libraries have a fixed lysine residue at the seventh position: lane 1, unmodified lysine; lane 2, AcK; lane 3, SuccK. (b) Western blot analysis of recombinant E. coli isocitrate dehydrogenase (Icda), GADPH (GapA) and serine hydroxymethyltransferase (GlyA) competed with a lysine-succinylated (S) or an unmodified (U) peptide library. The same amounts of sample were loaded in both lanes for each protein. (c) Western blot analysis of lysine succinylation competed with a lysine succinylated (right) or an unmodified (left) peptide library in protein whole-cell lysates of E. coli (K-12 MG1655), S. cerevisiae (strain BY4741), D. melanogaster (S2 cells), M. musculus (C3H10 (T1/2) cells) and H. sapiens (Hela cells).