Histone H3.3 Protein (G34W, Nucleosome)

Catalog No. ABIN7938949

This Recombinant Histone H3.3 protein is expressed in Escherichia coli (E. coli).

Quick Overview for Histone H3.3 Protein (G34W, Nucleosome) (ABIN7938949)

Target: Histone H3.3 (H3F3A) (H3 Histone, Family 3A (H3F3A))

Protein Type: Recombinant

Origin: Human

Source: Escherichia coli (E. coli)

Application: Screening Assay (ScA), Enzyme Kinetics (EK)

Purity: >95 % pure by SDS-PAGE

Product Details

Protein Characteristics: G34W, Nucleosome

Purpose: Recombinant Polynucleosomes H3.3 (G34W)

Characteristics: Nucleosomes comprise the smallest subunit of chromatin, they consist of 147 bp of DNA wrapped around an octamer of core histone proteins (H2A, H2B, H3 and H4). Therefore, nucleosomes are more physiologically relevant substrates than histones and histone-derived peptides for use in 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.

Application Details

Application Notes: Recombinant Polynucleosomes H3.3 (G34W) are suitable as substrate for use in the study of enzyme kinetics, inhibitor screening, and selectivity profiling.

Restrictions: For Research Use only

Handling

Format: Liquid

Buffer: Human Recombinant Polynucleosomes H3.3 (G34W) (20 μg protein+ 24 μg DNA) are supplied in 10 mM Tris pH 8.0, 1 mM EDTA, 2 mM DTT, 20 % glycerol.

Preservative: Dithiothreitol (DTT)

Precaution of Use: This product contains Dithiothreitol (DTT): a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Handling Advice: Avoid repeated freeze/thaw cycles,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. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.

Target Details

Target: Histone H3.3 (H3F3A) (H3 Histone, Family 3A (H3F3A))

Alternative Name: Histone H3.3

Background:

Protein 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.

Short Description: Recombinant Polynucleosomes H3.3 (G34W), 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 Gly34Trp, 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.

Aliases: Histone H3, H3.3, Histone 3, G34W