Sequential Chromatin Immunoprecipitation Assay
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Corresponding Organization : University of Edinburgh
Other organizations : Institut de génétique et de biologie moléculaire et cellulaire, Inserm, Université de Strasbourg, Centre National de la Recherche Scientifique, Helmholtz Zentrum München
Variable analysis
- Antibodies recognizing H3K122ac and H3K64ac
- Chromatin fragments sheared to an average length of ~100 - 200 bp
- Chromatin immunoprecipitated using H3K4me3 and H3K27me3 antibodies
- Chromatin immunoprecipitated using H3K122ac antibody
- Chromatin quantified by qPCR and expressed as % of input bound
- MESCs cross-linked in 1% formaldehyde for 10 min
- Glycine added to final concentration of 0.125 M to quench cross-linking
- Roche Protease Inhibitor Cocktail used in RIPA buffer
- H3K4me3 antibody
- H3K27me3 antibody
- Not explicitly mentioned
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