Mononucleosomes H3.1 Recombinant Protein | H3.1 recombinant protein

Recombinant Mononucleosomes H3.1 (K9M)

Cat. Num. AAA389303

• Synonyms: Mononucleosomes H3.1; Recombinant Mononucleosomes H3.1 (K9M); H3.1 recombinant protein

• Host: E Coli
• Form/Format: Recombinant Mononucleosomes H3.1 (K9M)(20ug protein + 20ug DNA) is supplied in 10mM Tris-HCl pH8.0, 1mM EDTA, 2mM DTT, and 20% glycerol.

• Protein Species: Human
• Protein Details: Recombinant Mononucleosomes H3.1 (K9M) consist of a 167 bp of 601 DNA without tags 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), H3.1 that includes amino acids 1-136 (end)(accession number NM_003529) with a point mutation Lys9Met, 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 H3.1 (K9M) 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 H3.1 (K9M) DNA gel Recombinant Mononucleosomes H3.1 (K9M) 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 nucleosomes. Lane 3: Intact mononucleosomes H3.1 (K9M). Intact mononucleosomes H3.1 (K9M) 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 H3.1 (K9M) 12.5% SDS-PAGE gel stained with Coomassie Blue MW: 108 kDa Purity: >95%)

Western Blot (WB)

(Western blot assay for Recombinant Mononucleosomes H3.1 (K9M) 0.5 ug Recombinant Histone H3.1 (K9M) which was used for assembly of nucleosomes was incubated with 0 or 5 nM G9A respectively in reaction buffer containing 50 mM Tris-HCl pH 8.6, 0.02% Triton X-100, 2 mM MgCl2, 1 mM TCEP and 50 uM SAM for 3 hr at room temperature. Half of the reactions were run on a 12.5% SDS-PAGE gel and detected with H3K9me2 antibody . Histone H3.1 was used as a control. The Western Blot result shows that only the K9 of histone H3.1 can be methylated by G9A.)

Related Product Information for H3.1 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. 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. Mutations in histone H3 occur with high genetic penetrance within rare pediatric gliomas and sarcomas. In H3 variants, the mutation is most often a lysine-to-methionine (K-M) mutation, occasionally glycine mutations (G34R/V/W/L) occur too. According to researchers, mutations at H3K4: out of a total of 9 mutations at this site, 8 were a K4M/I substitution. More K-to-M/I mutations were observed, raising the possibility that the functional effects associated with known K-to-M/I changes (that is, function in a dominant fashion to block the methylation of corresponding lysines on wild type histones) may extend to additional contexts.

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. 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. Mutations in histone H3 occur with high genetic penetrance within rare paediatric gliomas and sarcomas. In H3 variants, the mutation is most often a lysine-to-methionine (K-M) mutation, occasionally glycine mutations (G34R/V/W/L) occur too. According to researchers, mutations at H3K4: out of a total of 9 mutations at this site, 8 were a K4M/I substitution. More K-to-M/I mutations were observed, raising the possibility that the functional effects associated with known K-to-M/I changes (that is, function in a dominant fashion to block the methylation of corresponding lysines on wild type histones) may extend to additional contexts.

Product Categories/Family for H3.1 recombinant protein

Recombinant Proteins; Histone

NCBI and Uniprot Product Information

• NCBI GenBank Nucleotide #: NM_003512
• Molecular Weight: Molecular weight of histone octamer is 108kDa