Histone 3 Protein (H3K27me3, Nucleosome, pseudo-methylated) (Biotin)

Catalog No. ABIN7939085

Recombinant Histone 3 protein expressed in Escherichia coli (E. coli).

Quick Overview for Histone 3 Protein (H3K27me3, Nucleosome, pseudo-methylated) (Biotin) (ABIN7939085)

Target: Histone 3 (H3) (Histone H3 (H3))

Protein Type: Recombinant

Origin: Human

Source: Escherichia coli (E. coli)

Product Details

Protein Characteristics: H3K27me3, Nucleosome, pseudo-methylated

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

Purpose: Recombinant Mononucleosomes H3K27me3 (MLA) - biotin

Characteristics: Recombinant Mononucleosomes H3K27me3 (MLA) - biotin consist of a 167 bp of 601 DNA with a 5'-biotin tag 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 trimethyl Lys27 (H3K27me3) (MLA) that includes amino acids 1-136 (end) (accession number NP_003520.1) with two substitutions (cysteine to serine at amino acid 96 and cysteine to alanine at amino acid 110), and H4 that includes amino acids 1-103 (end) (accession number NM_003548). All of these histones were expressed in E. coli. The molecular weight of histone octamer is 108 kDa. 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 histones are suitable for use as positive controls in the analysis of histone posttranslational modifications, as substrates for histone modification enzymes, or to generate chromatin in vitro.

Comment:

Pseudomethylated Lysines have been generated using the partented Methylated Lysin Analog (MLA) technology. In MLA, methylated histones are generated via a chemical alkylation reaction that substitutes a methylated analog of lysine, aminoethylcysteine, for the existing lysine at the desired residue. Aminoethylcysteine is structurally and chemically similar to lysine, though it contains a sulfide substitution in place of the lysine gamma-methylene. The MLA technique provides precise control over the site and degree of methylation.

Restrictions: For Research Use only

Handling

Format: Liquid

Buffer: Recombinant Mononucleosomes H3K27me3 (MLA) - biotin (20 μg protein + 20 μg DNA) are supplied 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 3 (H3) (Histone H3 (H3))

Alternative Name: Histone H3

Background:

Protein Background: Histone H3 is one of the core components of the nucleosome. The nucleosome is the smallest subunit of chromatin and consists of 146 base pairs of DNA wrapped around an octamer of core histone proteins (two each of H2A, H2B, H3 and H4). Histone H1 is a linker protein, present at the interface between the nucleosome core and DNA entry/exit points. Recombinant Histone H3K27me3 (MLA) has been generated using the patented Methylated Lysine Analog (MLA) technology. In MLA, methylated histones are generated via a chemical alkylation reaction that substitutes a methylated analog of lysine, aminoethylcysteine, for the existing lysine at the desired residue. Aminoethylcysteine is structurally and chemically similar to lysine, though it contains a sulfide substitution in place of the lysine g-methylene. The MLA technique provides precise control over the site and degree of methylation. The MLA technology is covered under U.S. Patent No. 8,278,112.

Short Description: Recombinant Mononucleosomes H3K27me3 (MLA) - biotin consist of a 167 bp of 601 DNA with a 5'-biotin tag 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 trimethyl Lys27 (H3K27me3) (MLA) that includes amino acids 1-136 (end) (accession number NP_003520.1) with two substitutions (cysteine to serine at amino acid 96 and cysteine to alanine at amino acid 110), 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.

Molecular Weight: 108 kDa

NCBI Accession: NP_003520