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Nucleosomes (AA 1-103), (AA 1-126), (AA 1-130), (AA 1-136), (biotinylated), (C-Term) protein (Biotin)

Spezies: Human Wirt: Escherichia coli (E. coli) Recombinant EAA, ScA
Produktnummer ABIN2669717
  • Target
    Nucleosomes
    Protein-Typ
    Recombinant
    Proteineigenschaft
    C-Term, AA 1-130, AA 1-126, AA 1-136, AA 1-103, biotinylated
    Spezies
    Human
    Quelle
    • 2
    Escherichia coli (E. coli)
    Aufreinigungstag / Konjugat
    Biotin
    Applikation
    Enzyme Activity Assay (EAA), Screening Assay (ScA)
    Produktmerkmale
    Recombinant Nucleosomes (H3.3) - biotinylated, Human consists of 5000 bp of DNA (plasmid) and two molecules each of histones H2A that includes amino acids 1-130 (end) (accession number NM_003512), H2B that includes amino acids 1-126 (end) (accession number NM_003518), H3.3 that includes amino acids 1-136 (end) (accession number NM_005324) with a C-terminal Biotin tag, and H4 that includes amino acids 1-103 (end) (accession number NM_003548). Recombinant Nucleosomes (H3.3) - biotinylated, Human was generated in E. coli cells and has an observed molecular weight of 110 kDa.
  • Applikationshinweise
    Recombinant Nucleosomes (H3.3) - biotinylated is suitable for use in the study of enzyme kinetics, inhibitor screening, and selectivity profiling. HMT Assay Conditions: 50 mM TrisCl, pH 8.6, 0.02 % Triton X-100, 2 mM MgCl2, 1 mM TCEP, 100 μM SAM, 30 ng/μl Recombinant Nucleosomes (H3.3) - biotinylated, 30 ng/μl DOT1L (1-416 aa) at 2 hours at room temperature. Activity was detected by fluorography.
    Beschränkungen
    Nur für Forschungszwecke einsetzbar
  • Konzentration
    0.85 μg/μL
    Handhabung
    Avoid repeated freeze/thaw cycles and keep on ice when not in storage.
    Lagerung
    -80 °C
    Informationen zur Lagerung
    Recombinant proteins in solution are temperature sensitive and must be stored at -80°C to prevent degradation.
  • Target
    Nucleosomes
    Hintergrund
    In vivo, histones are wrapped around by DNA in chromatin. Therefore, nucleosomes are more physiologically relevant substrates than histones and histone-derived peptides for in vitro studies. More importantly, some histone methyltransferases are significantly more active, as well as specific, when using nucleosomal substrates in HMT assays, such as DOT1L and NSD family enzymes. Nucleosomes are also widely used in histone methyltransferase screening assays to identify small molecular inhibitors for drug discovery, one of such examples is PRC2(EZH2). Histone H3.1 and Histone H3.3 are the two main Histone H3 variants found in plants and animals. They are known to be important for gene regulation. Histone H3.1 and H3.3 have been shown to demonstrate unique genomic localization patterns thought to be associated with their specific functions in regulation of gene activity. Specifically, Histone H3.1 localization is found to coincide with genomic regions containing chromatin repressive marks (H3K9me3, H3K27me3 and DNA methylation), whereas Histone H3.3 primarily colocalizes with marks associated with gene activation (H3K4me3, H2BK120ub1, and RNA pol II occupancy). Deposition of the Histone H3.1 variant into the nucleosome correlates with the canonical DNA synthesis-dependent deposition pathway, whereas Histone H3.3 primarily serves as the replacement Histone H3 variant outside of S-phase, such as during gene transcription. Aberrant localization of these variants is also known to correlate with certain cancers.
    Molekulargewicht
    110 kDa
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