• My NEB
  • Print
  • PDF
  • Histone H3.1/H4 Tetramer Human, Recombinant

    Description

    Histone H3.1 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. Histone H3.1, an H3 variant that has thus far only been found in mammals, is replication dependent and is associated with gene activation and gene silencing. The tetramer 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. Because the histones are folded with their subunit partners, the tetramer may be a better substrate for specific enzymes and modifications.

    Human Histone H3.1 Protein Sequence: ARTKQ TARKS TGGKA PRKQL ATKAA RKSAP ATGGV KKPHR YRPGT VALRE IRRYQ KSTEL LIRKL PFQRL VREIA QDFKT DLRFQ SSAVM ALQEA CEAYL VGLFE DTNLC AIHAK RVTIM PKDIQ LARRI RGERA (Genbank accession number: AAN10051)

    Human Histone H4 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 H3.1/H4 Tetramer Human, Recombinant
    SDS-PAGE analysis of Histone H3.1/H4 Tetramer Human, Recombinant
    Lane 1: NEB Protein Ladder (NEB #P7703 )
    Lane 2: Histone H3.1 (NEB #M2503)
    Lane 3: Histone H4 (NEB #M2504)
    Lane 4–6: 2.0, 4.0, 8.0 µg Histone H3.1/H4 Tetramer

    Product Source

    Purified H3.1 and H4 (NEB #M2503 and NEB #M2504) were denatured, refolded and the Histone H3/H4 Tetramer was purified by gel filtration.

    Properties and Usage

    Usage Concentration

    10 μM

    Storage Temperature

    -20°C

    Storage Conditions

    20 mM Tris-HCl
    2 M NaCl
    1 mM DTT
    1 mM EDTA
    pH 8.0 @ 25°C

    Notes

    1. 10 μM, 0.53 mg/ml is calculated using the molar extinction coefficient for Histone H3.1/H4 Tetramer (21,360) and its absorbance at 280 nm (5,6)
    2. To use as a substrate in an enzyme modification assay or other salt sensitive protocol, use 1 to 2 µl of the dimer in a minimum 20 µl reaction so that the salt concentration in the reaction is ≤ 200 mM.
    3. Because of high salt, the solution does not always remain frozen.

    References

    1. Kornberg, R.D. (1977). Annu. Rev. Biochem,. 46, 931-954.
    2. Van Holde, K.E. (1989). Chromatin,. 1-97.
    3. Hake, S.B. et al. (2006). J. Biol. Chem,. 281, 559-568.
    4. Mersha, F unpublished observation.
    5. Pace, C.N. et al. (1995). Protein Science. 4, 2411-2423.
    6. Gill, S.C. and von Hippel, P.H. (1989). Anal. Biochem. 182, 319-326.

    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.

    Interactive Tools

    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.
    • 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.

    Safety Data Sheet

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

    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.