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Sirtuins

Volume 1077 of the series Methods in Molecular Biology pp 105-120

Date:

SILAC-Based Quantification of Sirt1-Responsive Lysine Acetylome

  • Yue ChenAffiliated withBen May Department for Cancer Research, University of Chicago
  • , Gozde ColakAffiliated withBen May Department for Cancer Research, University of Chicago
  • , Yingming ZhaoAffiliated withBen May Department for Cancer Research, University of Chicago

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Abstract

Stable Isotope Labeling by Amino acids in Cell culture (SILAC) is one of the in vivo metabolic labeling methods widely used for dynamic analysis of protein modifications. Here, we describe a general approach to applying SILAC, in combination with affinity enrichment of acetyllysine peptides and mass spectrometry, to study the dynamic changes of the Lysine acetylome in response to Sirt1. The method should be applicable to quantify changes to other post translational modifications in diverse cellular systems.

Key words

SILAC Quantification Sirt1 Immunoaffinity purification Lysine acetylation Nano HPLC mass spectrometry