Recombinant Histone H3K27me3 (MLA)

Aliases: Histone H3, H3, MLA
Catalog No: 31579 Format: 100 µg $320 Buy
Catalog No: 31979 Format: 1 mg $935 Buy

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Expressed In: E. coli Protein Species: Human

Methylated Lysine Analog

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 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 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 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 modifiations, as substrates for histone modifiation enzymes, or to generate chromatin in vitro.

Protein Details

Recombinant human Histone H3 trimethyl Lys27 (H3K27me3) (accession number NP_003520.1) was expressed in E. coli cells and contains two substitutions (cysteine to serine at amino acid 96 and cysteine to alanine at amino acid 110). Recombinant methylated histones are specifically methylated via a chemical alkylation reaction that introduces a methyl lysine analog (MLA). The molecular weight of the protein is 15,285 Daltons. The recombinant histone is >92% pure by SDS-PAGE.
Protein Sequence (a.a. substitutions in italics): ARTKQTARKS TGGKAPRKQL ATKAARCSAP ATGGVKKPHR YRPGTVALRE IRRYQKSTEL LIRKLPFQRL VREIAQDFKT DLRFQSSAVM ALQEASEAYL VGLFEDTNLA AIHAKRVTIM PKDIQLARRI RGERA

References

This product was used in the following publications:
Nucleic Acids Res. (2019). Jun 5. pii: gkz493. PMID: 31165872. (Histone Demethylation Assay)

 

Recombinant Histone H3K27me3 (MLA) gel.
Histone H3K27me3 (MLA) was run on a 12.5% SDS- PAGE gel and stained with Coomassie Blue.

ESI-TOF Mass Spec analysis of Recombinant Histone H3K27me3 (MLA)
Expected mass: 15285 Da
Observed mass: 15285 Da

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.