Product Class: Kit

EpiMark® Nucleosome Assembly Kit

  • Catalog # E5350 was discontinued on December 18, 2023

Product Introduction

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 
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Catalog # Size Concentration

Product Information

Description

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.


This product is related to the following categories:

Kit Components

Kit Components

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

Properties & Usage

Application Features

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

References

  1. Kornberg, R.D. (1977). Annu. Rev. Biochem . 46, 931-954.
  2. van Holde, K.E. (1989). Chromatin. 1-497.
  3. Li, Y et al. (2009). Jour. of Biol. Chem. 284, 34283-34295.
  4. Qi, Hank H. et al. (2010). Nature. 466, 503-507.
  5. Luger et al. (2004). Methods in Enzymology. 375, 23-62.

Protocols, Manuals & Usage

Protocols

  1. Introduction (E5350)
  2. Dilution Assembly Protocol (E5350)
  3. Dialysis Assembly Protocol (E5350)
  4. Gel Shift Assay for EpiMark™ Nucleosome Assembly Kit (E5350)
  5. DNA Concentration Formulas for EpiMark™ Nucleosome Assembly Kit (E5350)

Manuals

The Product Manual includes details for how to use the product, as well as details of its formulation and quality controls.

FAQs & Troubleshooting

FAQs

  1. Where can I find more information for specific proteins and DNA found in the kit?
  2. What is the sequence of the Nucleosome Control DNA included in the kit?
  3. How much Nucleosome Control DNA is provided in the kit?
  4. Are the components of the kit available for purchase separately?

Tech Tips

It is critical to always add the Dimer and Tetramer last when preparing the assembly reaction.

Quality, Safety & Legal

Quality Assurance Statement

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]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

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.

Trademarks

EPIMARK® is a registered trademark of New England Biolabs, Inc.
TRIDYE™ is a trademark of New England Biolabs, Inc.

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