Histone H3.3 Protein (K27M, Nucleosome) (Biotin)

Catalog No. ABIN7938944

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

Quick Overview for Histone H3.3 Protein (K27M, Nucleosome) (Biotin) (ABIN7938944)

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)

Product Details

Protein Characteristics: K27M, Nucleosome

Purification tag / Conjugate: This Histone H3.3 protein is labelled with Biotin.

Purpose: Recombinant Mononucleosomes H3.3 (K27M) - biotin

Characteristics: Recombinant Mononucleosomes H3.3 (K27M) - biotinylated, consist of a 167 bp of 601 DNA with 5' biotin tag and two molecules each of histones H2A that includes amino acids 1-130 (end) (accession number NP_003503.1), H2B that includes amino acids 1-126 (end) (accession number NP_003509.1), H3.3 that includes amino acids 1-136 (end) (accession number NP_002098.1) with a point mutation Lys27Met, and H4 that includes amino acids 1-103 (end) (accession number NP_003539.1). All of these histones were expressed in E. coli. The molecular weight of histone octamer is ~108 kDa. The recombinant protein is ≥95 % by SDS-PAGE gel. 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 Mononucleosomes H3.3 (K27M) - biotinylated are suitable for use in the study of enzyme kinetics, inhibitor screening, and selectivity profiling.

Restrictions: For Research Use only

Handling

Buffer: Recombinant Mononucleosomes H3.3 (K27M) - biotinylated (20 μg protein + 20 μg DNA) is supplied at a protein concentration of 0.75 μg/ul in 10 mM Tris-HCl, pH 8.0, 1 mM EDTA, 2 mM DTT and 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 consists 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, mono-ubiquitination, tec. 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 were involved in multiple biology 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 seen in chondroblastomas and mutation to G34W/L in giant cell tumors of bone, which are diseases of adolescents and young adults. Histone H3.3 mutations may 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 Mononucleosomes H3.3 (K27M) - biotinylated, consist of a 167 bp of 601 DNA with 5' biotin tag and two molecules each of histones H2A that includes amino acids 1-130 (end) (accession number NP_003503.1), H2B that includes amino acids 1-126 (end) (accession number NP_003509.1), H3.3 that includes amino acids 1-136 (end) (accession number NP_002098.1) with a point mutation Lys27Met, and H4 that includes amino acids 1-103 (end) (accession number NP_003539.1). All of these histones were expressed in E. coli cells. The molecular weight of histone octamer is ~108 kDa. The recombinant protein is ≥95 % by SDS-PAGE gel.

Molecular Weight: 108 kDa

NCBI Accession: NP_003503