Recombinant Mononucleosomes H3K36me3 (MLA) - biotin

Catalog No: 81076 Format: 20 µg $450 Buy
Catalog No: 81776 Format: 1 mg $5,845 Buy

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Expressed In: E. coli Protein Species: Human

Methylated Lysine Analog

Recombinant Mononucleosomes H3K36me3 (MLA) - biotin 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 ϒ-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.

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

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 H3K36me3 (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 ϒ-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. 

Application Notes

Recombinant histones are suitable for use as positive controls in the analysis of histone post-translational modifications, as substrates for histone modification enzymes, or to generate chromatin in vitro.

Protein Details

Mononucleosomes H3K36me3 (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 Lys36 (H3K36me3) (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. 

 

Recombinant Mononucleosomes H3K36me3 (MLA) - biotin SDS-PAGE gel

13% SDS-PAGE Coomassie staining

MW: 108 kDa

Purity: >95% 

Recombinant Mononucleosomes H3K36me3 (MLA) - biotin agarose gel
Recombinant Mononucleosomes H3K36me3 (MLA) - biotin were run on a 2% agarose gel and stained with ethidium bromide. Lane 1: DNA marker. Lane 2: 601 DNA. Lane 3: Intact monnucleosomes H3K36me3. Intact mononucleosomes H3K36me3 migrated much higher than free 601 DNA. The agarose gel shows that almost all of 601 DNA wrap histone octamers to form nucleosomes. 

Western Blot analysis for Recombinant Mono-nucleosomes H3K36me3 (MLA) – biotin
Recombinant Mononucleosomes H3K36me3 (MLA) - biotin (Lane 1) and unmodified nucleosomes (Lane 2) were detected with anti-H3K36me3 antibody and anti-H4 antibody, respectively. H4 was detected as loading control. Only Recombinant Mononucleosomes H3K36me3 (MLA) - biotin can be detected by H3K36me3 antibody.

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 for research use only and is not for use in diagnostic procedures. This product is guaranteed for 6 months from date of arrival.