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  • Isolation and characterization of histone H3 lysine 4 demethylase-containing complexes.

Isolation and characterization of histone H3 lysine 4 demethylase-containing complexes.

Methods (San Diego, Calif.) (2006-11-15)
Min Gyu Lee, Christopher Wynder, Jessica Norman, Ramin Shiekhattar
ABSTRACT

Histone methylation is involved in the regulation of many cellular processes. In the past 2 years, several histone demethylases including BHC110/LSD1 have been characterized. BHC110, the first known histone lysine demethylase, removes methyl groups from methylated histone H3 lysine 4 and has been found in many multi-protein complexes. Using one-step affinity purification, we have isolated enzymatically active BHC110-containing complexes. Here, we detail the methods used for the isolation and characterization of these histone demethylase complexes from a human stable cell line.

MATERIALS
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Product Description

Sigma-Aldrich
Monoclonal ANTI-FLAG® M2 antibody produced in mouse, clone M2, purified immunoglobulin (Purified IgG1 subclass), buffered aqueous solution (10 mM sodium phosphate, 150 mM NaCl, pH 7.4, containing 0.02% sodium azide)