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  • Histone H4 Human, Recombinant

    Description

    Histone H3 combines with Histone H4 to form the H3/H4 tetramer. Two H2A/H2B heterodimers interact with an H3/H4 tetramer to form the histone octamer (1,2). Histone H4 is also modified by various enzymes and can act as a substrate for them. These modifications have been shown to be important in gene regulation.

    Protein Sequence:
    SGRGK GGKGL GKGGA KRHRK VLRDN IQGIT KPAIR RLARR GGVKR ISGLI YEETR GVLKV FLENV IRDAV TYTEH AKRKT VTAMD VVYAL KRQGR TLYGF GG (Genbank accession number: AAM83108)

    SDS-PAGE analysis of Histone H4 Human, Recombinant
    Lane 1 & 7: NEB Protein Ladder (NEB #P7703),
    Lane 2 thru 6: 0.5–10.0 µg Histone H4 Human, Recombinant.

    ESI-TOF Analysis of Histone H4 Human, Recombinant.

    Properties and Usage

    Storage Temperature

    -20°C

    Storage Conditions

    300 mM NaCl
    1 mM EDTA
    20 mM sodium phosphate
    pH 7.0 @ 25°C

    Quality Control

    Quality Control Assays

    The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page.
    • Endonuclease Activity (Nicking):
      The product is tested in a reaction containing a supercoiled DNA substrate. After incubation for 16 hours the percent converted to the nicked form is determined by agarose gel electrophoresis.
    • Exonuclease Activity (Radioactivity Release):
      The product is tested in a reaction containing a radiolabeled mixture of single and double-stranded DNA. After incubation for 4 hours the exonuclease activity is determined by the % release of radioactive nucleotides.
    • Histone Modification (Radioactivity Incorporation):
      The Histone is tested in a reaction containing a specific Methyltransferase and radiolabeled SAM. After incubation the number of pmoles of methyl group per unit of Methyltransferase transferred to the Histone is determined by radioactive incorporation.
    • Molecular Weight Determination (Mass Spectrometry) :
      The molecular weight of the product is determined using mass spectrometry.
    • Protease Activity (SDS-PAGE):

      The product is tested for protease activity by incubation with a standard mixture of proteins resulting in no detectable degradation of the proteins as determined by SDS-PAGE.

    Notes

    1. The protein concentration (1 mg/ml, 89 µM) is calculated using the molar extinction coefficient for Histone H4 (5120) and its absorbance at 280 nm (3,4).  1.0 A280 units = 2.2 mg/ml.

    References

    1. Kornberg, R.D. (1977). Annu. Rev. Biochem. 46, 931-954.
    2. van Holde, K.E. (1989). Chromatin. 1
    3. Gill, S.C. and von Hippel, P.H. (1989). Anal. Biochem.. 182, 319-326.
    4. Pace, C.N. et al. (1995). Protein Science. 4, 2411-2423.

    Supporting Documents

    Material Safety Datasheets

    The following is a list of Material Safety Data Sheets (MSDS) that apply to this product to help you use it safely. The following file naming structure is used to name these document files: [Product Name] MSDS. For international versions please contact us at info@neb.com.

    Datacards

    The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.
    1. Do the histones need to be reconstituted?
    2. What are the recommended histone storage conditions?
    3. Are the histones fusion proteins or tagged proteins?
    4. Can the histones be used as substrates for protein modification enzymes? Which ones?
    After thawing on ice, mix well by pipetting the solution up and down. Do not centrifuge.