The EpiMark™ Nucleosome Assembly Kit is used to make unmodified recombinant human nucleosomes with user-supplied DNA or the provided control DNA. Purified recombinant human Histone H2A/H2B Dimer and Histone H3.1/H4 Tetramer are mixed with DNA at 2 M NaCl. To generate nucleosomes, the salt concentration is lowered by dilution or dialysis allowing each histone tetramer to associate with two histone dimers and form the histone octamer on the DNA. A method for assaying nucleosome formation by gel shift assay is included. These nucleosomes may serve as a better substrate for enzymes that are inactive on the DNA or one of the core histones alone. Each reaction creates nucleosomes from ~1 µg of 208 bp DNA and may be scaled depending on the experiment.
Applications:
- ChIP assay
- HAT assay
- Enzymatic modification assay, (e.g., methylation)
Advantages:
- Highly pure, recombinant system
- Components stable for one year
- Pre-formed histone dimer and tetramer complexes simplify octamer formation
- Dilution protocol only requires a few hours for assembly
Kit Components:
- Histone H2A/H2B Dimer Human, Recombinant
- Histone H3.1/H4 Tetramer Human, Recombinant
- Nucleosome Control DNA