Enrichment and identification of acetylated and succinylated peptide assay
This protocol is extracted from research article:
Protein lysine de-2-hydroxyisobutyrylation by CobB in prokaryotes
Sci Adv, Jul 17, 2019; DOI: 10.1126/sciadv.aaw6703

For enrichment and identification of acetylation, we used the previously reported method similar to that for 2-hydroxyisobutyrylation (3). Briefly, after whole-cell proteins were digested, tryptic peptides were fractionated using high-pH reversed-phase HPLC, and then acetylated peptides were enriched using agarose-conjugated anti-acetyl lysine antibody. Each sample of enriched Kac peptides was analyzed by HPLC-MS/MS, and the final result was verified using MaxQuant software (v1.5.2.8). In contrast to acetylation assay, the samples used for succinylation were not fractionated by high-pH reversed-phase HPLC.

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