EpiMark® Nucleosome Assembly Kit |NEB酶试剂 New England Biolabs

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产品信息

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 AssemblyEpiMark® Nucleosome Assembly Kit |
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.

产品类别:
Discontinued (<3 years)

  • 优势和特性

    应用特性

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

  • 相关产品

    单独销售的组分

    • 核小体对照 DNA
    • 重组人源二聚体组蛋白 H2A/H2B
    • 重组人源四聚体组蛋白 H3.1/H4

  • 参考文献

    1. Kornberg, R.D. (1977). Annu. Rev. Biochem . 46, 931-954. PubMedID: 332067
    2. van Holde, K.E. (1989). Chromatin. 1-497.
    3. Li, Y et al. (2009). Jour. of Biol. Chem. 284, 34283-34295. PubMedID: 19808676
    4. Qi, Hank H. et al. (2010). Nature. 466, 503-507. PubMedID: 20622853
    5. Luger et al. (2004). Methods in Enzymology. 375, 23-62. PubMedID: 14870657

操作说明、说明书 & 用法

  • 操作说明

    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)

  • 说明书

    产品说明书包含产品使用的详细信息、产品配方和质控分析。

    • manualE5350

FAQs & 问题解决指南

  • 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?

  • 实验技巧

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