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  • Extraction, Identification, and Quantification of Histones from Small Quantities of Specific Brain Tissue.

Extraction, Identification, and Quantification of Histones from Small Quantities of Specific Brain Tissue.

Current protocols in neuroscience (2016-07-02)
Hassiba Beldjoud, Fany Messanvi, Nael Nadif Kasri, Benno Roozendaal
ABSTRACT

Histone posttranslational modifications (PTMs), by their action on the chromatin state, play a central role in the regulation of gene expression. The discovery that some PTMs in the brain are dynamically regulated by experience and environmental factors makes them an important subject for the study of plasticity changes in learning and memory, addiction, and psychiatric disorders. Current histone isolation protocols, however, require large amounts of tissue, which limits their application for analyzing small tissue samples from a specific brain region. We describe here a step-by-step protocol for histone extraction and isolation from 1 mm(3) of tissue from brain punches, which allows reproducible and reliable results for histone PTM identification and quantification without losing anatomical precision. © 2016 by John Wiley & Sons, Inc.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-Histone H3 Antibody, CT, pan, serum, Upstate®
Sigma-Aldrich
ChIPAb+ Trimethyl-Histone H3 (Lys27) - ChIP Validated Antibody and Primer Set, from rabbit, purified by using Protein A
Sigma-Aldrich
Anti-acetyl-Histone H2B Antibody, Upstate®, from rabbit
Sigma-Aldrich
Anti-phospho-Histone H3 (Ser10) Antibody, clone RR002, clone RR002, Upstate®, from mouse
Sigma-Aldrich
Anti-Histone H4 Antibody, pan, clone 62-141-13, rabbit monoclonal, clone 62-141-13, Upstate®, from rabbit
Sigma-Aldrich
Anti-trimethyl-Histone H3 (Lys4) Antibody, Upstate®, from rabbit
Roche
cOmplete, Mini Protease Inhibitor Cocktail, Tablets provided in EASYpacks
Sigma-Aldrich
Anti-acetyl-Histone H3 Antibody, from rabbit
Sigma-Aldrich
Sodium butyrate, ≥98.5% (GC)