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  • Histone H3.2 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. It 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. Histone H3.2, an H3 variant that is found in all eukaryotes except budding yeast, is replication dependent and is associated with gene silencing.

    Protein Sequence: ARTKQ TARKS TGGKA PRKQL ATKAA RKSAP ATGGV KKPHR YRPGT VALRE IRRYQ KSTEL LIRKL PFQRL VREIA QDFKT DLRFQ SSAVM ALQEA SEAYL VGLFE DTNLC AIHAK RVTIM PKDIQ LARRI RGERA (Genbank accession number: Q71DI3)

    ESI-TOF Analysis of Histone H3.2 Human, Recombinant

    H3.2 SDS-PAGE analysis of Histone H3.2 Human, Recombinant
    Lane 1 & 7:  NEB Protein Ladder (NEB #P7703).  
    Lane 2 through 6:  0.5, 1.0, 2.0, 5.0, 10.0 μg Histone H3.2 Human, Recombinant

    Product Source

    An E.coli strain that carries a plasmid encoding the cloned human histone H3.2 gene, HIST2H3A or HIST2H3C. (Genbank accession number: BC130637)

    Properties and Usage

    Storage Temperature

    -20°C

    Storage Conditions

    20 mM sodium phosphate
    300 mM (NH4)2SO4
    1 mM (NH4)2SO4
    1 mM DTT
    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 4 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.
    • N-terminal Protein Sequencing :
      Protein identity is confirmed using Edman Degradation to sequence the N-terminus of the intact protein.
    • 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. 1 mg/ml, 66 µM is calculated using the molar extinction coefficient for Histone H3.2 (3960) and its absorbance at 280 nm (4,5). 1.0 A280 units = 3.9 mg/ml
    2. Synonym: Histone H3/m, H3/o
    3. Gene Synonym: H3F2, H3FM

    References

    1. Kornberg, R.D. (1977). Annu. Rev. Biochem.. 46, 931-954.
    2. van Holde, K.E. (1989). Chromatin. 1-497.
    3. Hake, S.B. et al. (2006). J. Biol. Chem.. 281, 559-568.
    4. Gill, S.C. and von Hippel, P.H. (1989). Anal. Biochem.. 182, 319-326.
    5. 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. Are the histones fusion proteins or tagged proteins?
    2. Do the histones need to be reconstituted?
    3. What are the recommended histone storage conditions?
    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.