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

Recombinant Mononucleosomes H3.3 (R8C)

Cat. Num. AAA389318

• Gene Names: H3F3A; H3F3; H3.3A
• Synonyms: Mononucleosomes H3.3; Recombinant Mononucleosomes H3.3 (R8C); H3.3 recombinant protein

• Host: E Coli
• Form/Format: Recombinant Mononucleosomes H3.3 (R8C)(20ug protein + 20ug DNA) are supplied in 10mM Tris-HCl pH8.0, 1mM EDTA, 2mM DTT, and 20% glycerol.

• Protein Species: Human
• Protein Details: Recombinant Mononucleosomes H3.3 (R8C) consist of a 167 bp of 601 DNA 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 NP_002098.1) with a point mutation Arg8Cys, and H4 that includes amino acids 1-103 (end)(accession number NM_003548). All of these histones were expressed in E Coli cells.
• Notes: Recombinant Mononucleosomes H3.3 (R8C) is suitable for use as substrate for histone modification enzymes, or to generate chromatin in vitro.
• 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

Testing Data

(Recombinant Mononucleosomes H3.3 (R8C) DNA gel Recombinant Mononucleosomes H3.3 (R8C) were run on a 2% agarose gel and stained with ethidium bromide. Lane 1: DNA marker. Lane 2: 601 DNA which was used for assembly of nucleosome. Lane 3: Intact mononucleosomes H3.3 (R8C). Intact mononucleosomes H3.3 (R8C) migrated much higher than free 601 DNA. The agarose gel shows that almost all of 601 DNA wrapped histone octamers to form nucleosomes.)

SDS-Page

(Recombinant Mononucleosomes H3.3 (R8C) 12.5% SDS-PAGE gel with Coomassie Blue staining MW: 108 kDa Purity: >95%)

Related Product Information for H3.3 recombinant protein

Short Description: 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. 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 H3.3 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. Histones are linked to tumorigenesis primarily through alterations in their PTMs and the enzymes regulating these modifications, suggesting that they might disrupt the reading, writing, and/or erasing of these marks. Except for being near K9, which can be methylated or acetylated, R8 site itself can be methylated too. Mutations in histone H3 occur with high genetic penetrance within rare pediatric gliomas and sarcomas. Researchers found that mutations at H3 N-terminal residues at or near PTM sites including R2, R8, K18 and R26 might act as oncohistones.

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 signifcantly more active, as well as specifc, 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. 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 H3.3 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. Histones are linked to tumorigenesis primarily through alterations in their PTMs and the enzymes regulating these modifications, suggesting that they might disrupt the reading, writing, and/or erasing of these marks. Except for being near K9, which can be methylated or acetylated, R8 site itself can be methylated too. Mutations in histone H3 occur with high genetic penetrance within rare paediatric gliomas and sarcomas. Researchers found that mutations at H3 N-terminal residues at or near PTM sites including R2, R8, K18 and R26 might act as oncohistones.

Product Categories/Family for H3.3 recombinant protein

Recombinant Proteins; Histone

NCBI and Uniprot Product Information

• NCBI GI #: 4504279
• NCBI GeneID: 3020
• NCBI Accession #: NP_002098.1
• NCBI GenBank Nucleotide #: NM_003512
• UniProt Accession #: P84243; P06351; P33155; Q5VV55; Q5VV56; Q66I33; Q9V3W4
• Molecular Weight: 15,328 Da
• NCBI Official Full Name: histone H3.3
• NCBI Official Synonym Full Names: H3 histone, family 3A
• NCBI Official Symbol: H3F3A
• NCBI Official Synonym Symbols: H3F3; H3.3A
• NCBI Protein Information: histone H3.3
• UniProt Protein Name: Histone H3.3
• UniProt Gene Name: H3F3A
• UniProt Synonym Gene Names: H3.3A; H3F3
• UniProt Entry Name: H33_HUMAN

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 contains introns and its mRNA is polyadenylated, unlike most histone genes. The protein encoded is a replication-independent member of the histone H3 family. [provided by RefSeq, Jul 2008]

Uniprot Description

H3F3A: Variant histone H3 which replaces conventional H3 in a wide range of nucleosomes in active genes. Constitutes the predominant form of histone H3 in non-dividing cells and is incorporated into chromatin independently of DNA synthesis. Deposited at sites of nucleosomal displacement throughout transcribed genes, suggesting that it represents an epigenetic imprint of transcriptionally active chromatin. 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. The nucleosome is a histone octamer containing two molecules each of H2A, H2B, H3 and H4 assembled in one H3-H4 heterotetramer and two H2A-H2B heterodimers. The octamer wraps approximately 147 bp of DNA. Interacts with HIRA, a chaperone required for its incorporation into nucleosomes. Belongs to the histone H3 family.

Protein type: DNA-binding

Chromosomal Location of Human Ortholog: 1q42.12

Cellular Component: nucleoplasm; nuclear chromosome; protein complex; extracellular region; nucleosome; nucleus

Molecular Function: protein binding; nucleosomal DNA binding; protein heterodimerization activity

Biological Process: chromatin silencing at rDNA; nucleosome assembly; DNA replication-independent nucleosome assembly; negative regulation of gene expression, epigenetic; gene expression; blood coagulation; DNA methylation on cytosine; positive regulation of cell growth; regulation of gene expression, epigenetic