Cat.No.: | NUC-0100 |
Product Name: | Recombinant Polynucleosomes H3.3 (G34V) |
Product Overview: | Recombinant Polynucleosomes H3.3 (G34V), Human, consist of 5000 bp of DNA (plasmid pG5E4) 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 point mutation Gly34Val, and H4 that includes amino acids 1-103 (end) (accession number NM_003548). Plasmid pG5E4 contains 9 of 5S rDNA nucleosome positioning sequences of L. variegatus, 5 of GAL4 binding sites and E4 promoter. Every 5S rDNA can wrap one histone octamer to form a nucleosome. 5 of GAL4 binding sites and E4 promoter can wrap histone octamers to form dinucleosomes. A pG5E4 plasmid can warp histone octamers to form 11 nucleosomes. The recombinant protein is >95% pure by SDS-PAGE. |
Background: | In vivo, the nucleosome is the basic structural unit of chromatin. It is comprised of about 146 bp of DNA wrapped around a core of eight histones of four different types: H2A, H2B, H3 and H4. Histones are subject to posttranslational modifications, such as methylation, acetylation, phosphorylation and mono-ubiquitination. Histone modifications influence multiple chromatin-templated processes such as gene transcription, DNA repair and recombination. Besides the “major“ histones, there are some histone variants in specific regions of chromatin or in specific cell types. Histone variants are involved in multiple processes including chromosome segregation, DNA repair, transcriptional regulation and mRNA processing. Histone H3.3 point mutations (K27 and G34) are found in 1/3 of pediatric glioblastomas. Up to 78% of diffuse intrinsic pontine gliomas (DIPGs) carry K27M and 36% of non-brainstem gliomas carry either K27M or G34R/V mutations. High- frequency mutation of histone H3 to K36M is observed in chondroblastomas, as is the mutation G34W/L in giant cell tumors of bone, diseases of adolescents and young adults. Histone H3.3 mutations drive pediatric glioblastoma through upregulation of MYCN. 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. |
Applications: | Suitable for use in enzyme kinetics, inhibitor screening, and selectivity profiling. |
Storage: | Store at 4℃ if entire vial will be used within 2-4 weeks. Store at -20℃ or -80℃ for longer periods of time. Avoid multiple freeze-thaw cycles. |
Appearance: | Liquid |
Species: | Human |
Formulation: | 10 mM Tris pH 8.0, 1 mM EDTA, 2 mM DTT, 20% glycerol. |
Expression System: | E. coli |
Warning: | This product is for research use only. Not for use in diagnostic or therapeutic procedures. |
Product Types | ||
◆ Nucleosomes | ||
NUC-0007 | Recombinant Tetranucleosomes H3.3 | Inquiry |
NUC-0008 | Recombinant Mononucleosomes H3.3 | Inquiry |
◆ Synthetic Peptides | ||
SP-0009 | Histone H4 peptide (1-21), Biotin-labeled | Inquiry |
SP-0010 | Histone H4 peptide (1-21) | Inquiry |
SP-0012 | Histone H3 peptide (21-44), Biotin-labeled | Inquiry |
Related Gene / Proteins | |||
HIC1 | HIF1A | HIF1AN | HINFP |
HIPK1 | HIPK2 | HIRA | Histone |
Histone H1 | Histone H2A | Histone H2B | Histone H3 |
Histone H4 | HIV-1 reverse transcriptase |
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