New England Biolabs Canada
Expand menu
Sign In or Sign UpMy NEB

EpiMark® Nucleosome Assembly Kit

Catalog # Concentration Size List Price Quantity Your Price
E5350S 20 reactions
Please Inquire
Catalog # Size List Price Your Price
E5350S 20 reactions
Please Inquire
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

The Epimark Nucleosome Assembly Kit is used to make unmodified recombinant human nucleosomes with user-supplied DNA or provided control DNA.

  • Highly pure, recombinant proteins
  • Pre-formed histone dimer and tetramer complexes simplify octamer formation
  • Components stable for one year
  • Dilution protocol requires only a few hours for assembly
  • Applications include ChIP, HAT and enzymatic modification assays 
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 (1,2). A method for assaying nucleosome formation by gel shift assay is included. These nucleosomes may serve as better substrates for enzymes that are inactive on the DNA or one of the core histones alone (3,4). Each reaction creates nucleosomes from ~1 µg of 208 bp DNA and may be scaled depending on the experiment.

Nucleosome AssemblyNucleosome formation

A gel shift assay allows visualization of complex formation when various ratios of Octamer* to DNA are used to form mononucleosomes using the provided nucleosome control DNA. When assembly reactions are run on 6% polyacrylamide gel in 0.5X TBE, a characteristic shift occurs from 208 bp to approximately 700 bp.

  • M: Low Molecular Weight DNA Ladder (NEB #N3233)
  • Lane 1: Nucleosome Control DNA
  • Lane 2: 0.5:1 ratio of Octamer* to DNA
  • Lane 3: 1:1 ratio of Octamer* to DNA.

*Octamer = 2:1 mix of Histone H2A/H2B Dimer and Histone H3.1/H4 Tetramer.


Application Features
  • ChIP assay
  • HAT assay (Histone Acetyltransferase Assay)
  • Enzymatic modification assay, (e.g., methylation)

Properties & Usage



Materials Sold Separately
References
  • Kornberg, R.D. (1977). Annu. Rev. Biochem . 46, 931-954. PubMedID: 332067
  • van Holde, K.E. (1989). Chromatin. 1-497.
  • Li, Y et al. (2009). Jour. of Biol. Chem. 284, 34283-34295. PubMedID: 19808676
  • Qi, Hank H. et al. (2010). Nature. 466, 503-507. PubMedID: 20622853
  • Luger et al. (2004). Methods in Enzymology. 375, 23-62. PubMedID: 14870657
Quality Control Assay
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Specifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Legal And Disclaimer

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.EPIMARK® is a registered trademark of New England Biolabs, Inc.
TRIDYE™ is a trademark of New England Biolabs, Inc.

Other Products You May Be Interested In
 Top