Recombinant Mononucleosomes H3.1 (K18M)

Catalog No: 81255 Format: 20 µg $475 Buy

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Expressed In: E. coli Protein Species: Human

Contents

A representative Technical Data Sheet (TDS) is provided here. Please refer to the lot-specific TDS you will receive with your order for the lot-specific buffer contents and protein concentration.

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.

Application Notes

Recombinant Mononucleosomes H3.1 (K18M) is suitable for use as substrate for histone modification enzymes, or to generate chromatin in vitro.

Protein Details

Recombinant Mononucleosomes H3.1 (K18M) 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 Lys18Met, and H4 that includes amino acids 1-103 (end) (accession number NM_003548). All of these histones were expressed in E. coli cells. The molecular weight of histone octamer is 108 kDa.

 

Recombinant Mononucleosomes H3.1 (K18M) DNA gel
Recombinant Mononucleosomes H3.1 (K18M) 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 (K18M). Intact mononucleosomes H3.1 (K18M) migrated much higher than free 601 DNA. The agarose gel shows that almost all of 601 DNA wrapped histone octamers to form nucleosomes.

Recombinant Mononucleosomes H3.1 (K18M)
12.5% SDS-PAGE Coomassie staining
MW: 108 kDa
Purity: >90%

Storage

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.

Guarantee

This product is guaranteed for 6 months from date of receipt.

This product is for research use only and is not for use in diagnostic