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

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Protein Name
Protein Characteristics
C-Term, AA 1-130, AA 1-126, AA 1-136, AA 1-103, biotinylated
Escherichia coli (E. coli)
Protein Type
Purification tag / Conjugate
Enzyme Activity Assay (EAA), Screening Assay (ScA)
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Characteristics 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.
Background 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.
Molecular Weight 110 kDa
Application Notes 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.
Restrictions For Research Use only
Concentration 0.85 μg/μL
Handling Advice Avoid repeated freeze/thaw cycles and keep on ice when not in storage.
Storage -80 °C
Storage Comment Recombinant proteins in solution are temperature sensitive and must be stored at -80°C to prevent degradation.
Supplier Images
Western Blotting (WB) image for Nucleosomes (C-Term), (AA 1-130), (AA 1-126), (AA 1-136), (AA 1-103), (biotinylated) protein (Biotin) (ABIN2669717) DOT1L Activity assay using Recombinant Nucleosomes (H3.3) - biotinylated as substrate...
Western Blotting (WB) image for Nucleosomes (C-Term), (AA 1-130), (AA 1-126), (AA 1-136), (AA 1-103), (biotinylated) protein (Biotin) (ABIN2669717) Recombinant Nucleosomes (H3.3) - biotinylated protein gel. Recombinant Nucleosomes (H...
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