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Related Concept Videos

Chromatin Immunoprecipitation- ChIP02:36

Chromatin Immunoprecipitation- ChIP

Chromatin immunoprecipitation, or ChIP, is an antibody-based technique used to identify sites on DNA that bind to transcription factors of interest or histone proteins. It also helps determine the type of histone modifications such as acetylation, phosphorylation, or methylation.
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The histone proteins in the nucleosomes are post-translationally modified (PTM) to increase or decrease access to DNA. The commonly observed PTMs are methylation, acetylation, phosphorylation, and ubiquitination of lysine amino acids in the histone H3 tail region. These histone modifications have specific meaning for the cell. Hence, they are called "histone code". The protein complex involved in histone modification is termed as "reader-writer" complex.
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Related Experiment Video

Updated: May 8, 2026

Deacetylation Assays to Unravel the Interplay between Sirtuins (SIRT2) and Specific Protein-substrates
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Deacetylation Assays to Unravel the Interplay between Sirtuins (SIRT2) and Specific Protein-substrates

Published on: February 27, 2016

Assaying chromatin sirtuins.

Lei Zhong1, Barbara Martinez-Pastor1, Dafne M Silberman1

  • 1The Massachusetts General Hospital Cancer Center, Harvard Medical School, Boston MA 02114 USA.

Methods in Molecular Biology (Clifton, N.J.)
|September 10, 2013
PubMed
Summary

This study presents chromatin-based biochemical methods for identifying sirtuin substrates. These techniques help analyze sirtuin interactions with histones and chromatin proteins.

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Last Updated: May 8, 2026

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The ChroP Approach Combines ChIP and Mass Spectrometry to Dissect Locus-specific Proteomic Landscapes of Chromatin
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Published on: April 11, 2014

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Epigenetics

Background:

  • Sirtuins are a class of NAD+-dependent deacetylases and ADP-ribosyltransferases.
  • Nuclear sirtuin substrates primarily include histones and chromatin-associated proteins.
  • Understanding sirtuin-chromatin interactions is crucial for elucidating their biological roles.

Purpose of the Study:

  • To develop and present robust biochemical techniques for identifying sirtuin substrates within the chromatin context.
  • To facilitate the analysis of sirtuin biology related to chromatin modification and regulation.

Main Methods:

  • Development of chromatin immunoprecipitation (ChIP) protocols.
  • Implementation of acid-extraction protocols for analyzing chromatin-bound proteins.
  • Establishment of nucleosomal immunoprecipitation protocols to identify sirtuin chromatin interactors.

Main Results:

  • Detailed protocols for chromatin-based analysis of sirtuin substrates are provided.
  • The described methods enable the identification of proteins interacting with sirtuins on chromatin.
  • These techniques offer a framework for further investigation into sirtuin functions.

Conclusions:

  • The presented chromatin-based techniques are valuable tools for studying sirtuin-histone and sirtuin-chromatin interactions.
  • These methods enhance the biochemical analysis of sirtuin substrates in their native nuclear environment.
  • Further application of these protocols will advance the understanding of sirtuin-mediated epigenetic regulation.