Mononucleosomes H3.3 Recombinant Protein | H3.3 recombinant protein

Recombinant Mononucleosomes H3.3 (G34W)-biotin

Cat. Num. AAA389073

• Gene Names: HIST1H2AC; H2A/l; H2AFL; dJ221C16.4
• Purity: The purity ofThe recombinant protein is >=95% by SDS-PAGE analysis.
• Synonyms: Mononucleosomes H3.3; Recombinant Mononucleosomes H3.3 (G34W)-biotin; H3.3 recombinant protein

• Host: E Coli
• Purity/Purification: The purity ofThe recombinant protein is >=95% by SDS-PAGE analysis.
• Form/Format: Recombinant Mononucleosomes H3.3 (G34W)-biotinylated (20ug protein + 20ug DNA) is supplied at a protein concentration of 0.8ug/ul in 10mM Tris-HCl, pH8.0, 1mM EDTA, 2mM DTT and 20% glycerol.

• Protein Species: Human
• Conjugation: Biotinylated
• Notes: Recombinant Mononucleosomes H3.3 (G34W)-biotinylated are suitable for use in the study of enzyme kinetics, inhibitor screening, and selectivity profiling.
• Dry Ice Shipment: Extra charge fee may add to your shipping cost as dry ice is required to ship this product.
• Preparation and Storage: Recombinant proteins in solution are temperature sensitive and must be stored at -80 degree C to prevent degradation. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.; Shipping Temp: Dry Ice

SDS-Page

(Streptavidin pull-down for Recombinant Mononucleosomes H3.3 (G34W) – biotin. Recombinant Mononucleosomes H3.3 (G34W) - biotinylated were pulled down by streptavidin beads. Input nucleosomes (Lane 2) and the nucleosomes (Lane 3) pulled down by streptavidin were run on a 12.5% SDS-PAGE gel and stained with Coomassie blue. The SDS-PAGE gel result shows that almost all of biotinylated mononucleosomes H3.3 (G34W) are pulled down by streptavidin beads. * indicates streptavidin.)

Testing Data

(Recombinant Mononucleosomes H3.3 (G34W) – biotin, DNA gel. Recombinant Mononucleosomes H3.3 (G34W) - biotinylated were run on a 2% agarose gel and stained with ethidium bromide. Lane 1: DNA marker. Lane 2: 601 DNA. Lane 3: Intact monnucleosomes H3.3 (G34W). Intact mononucleosomes H3.3 (G34W) migrated much higher than free 601 DNA. The agarose gel result shows almost all of 601 DNA wrap histone octamers to form nucleosomes.)

Western Blot (WB)

(Histone methyltransferase activity assay comparing recombinant nucleosomes and histone octamers as substrates. 2 ug biotinylated mononucleosomes H3.3 (G34W) were incubated with 0.2 ug DOT1L (1-420) in 30 ul reaction system containing 50 mM Tris-HCl, pH 8.6, 0.02% Triton X-100, 2 mM MgCl2, 1 mM TCEP, 50 uM SAM for 3 h at room temperature. 6 ul reaction products were loaded and run on a 12.5% SDS-PAGE gel. Western Blot was used to detect the generation of reaction products (Anit-H3K79me1). DOT1L only was used as negative control.)

Related Product Information for H3.3 recombinant protein

Short Description: Recombinant Mononucleosomes H3.3 (G34W)-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 Gly34Trp, 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 the histone octamer is ~108 kDa. The purity of the recombinant protein is >=95% by SDS-PAGE analysis. 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.

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 mutations in histone H3 include K36M in chondroblastomas and 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.

Product Categories/Family for H3.3 recombinant protein

Recombinant Proteins; Histone

NCBI and Uniprot Product Information

• NCBI GI #: 4504245
• NCBI GeneID: 8334
• NCBI Accession #: NP_003503.1; NP_003509.1; NP_002098.1; NP_003539.1
• NCBI GenBank Nucleotide #: NM_003512.3
• UniProt Accession #: Q93077; O00775; O00776; O00777; O00778; Q540R1; B2R4F7
• Molecular Weight: 14,105 Da
• NCBI Official Full Name: histone H2A type 1-C
• NCBI Official Synonym Full Names: histone cluster 1 H2A family member c
• NCBI Official Symbol: HIST1H2AC
• NCBI Official Synonym Symbols: H2A/l; H2AFL; dJ221C16.4
• NCBI Protein Information: histone H2A type 1-C
• UniProt Protein Name: Histone H2A type 1-C
• UniProt Gene Name: HIST1H2AC
• UniProt Synonym Gene Names: H2AFL

NCBI Description

Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form an octamer, around which approximately 146 bp of DNA is wrapped in repeating units, called nucleosomes. The linker histone, H1, interacts with linker DNA between nucleosomes and functions in the compaction of chromatin into higher order structures. This gene is intronless and encodes a replication-dependent histone that is a member of the histone H2A family. Transcripts from this gene lack polyA tails but instead contain a palindromic termination element. This gene is found in the large histone gene cluster on chromosome 6. [provided by RefSeq, Aug 2015]

Uniprot Description

Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.