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Recombinant Proteins & Enzymes

Recombinant Mononucleosomes H3K79me1 (MLA) - biotin

Catalog No: 81137 Format: 20 µg $415 Buy Now
Expressed In: E. coli Protein Species: Human

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Methylated Lysine Analog

Recombinant Mononucleosomes H3K79me1 (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.

for Recombinant Mononucleosomes H3K79me1 (MLA) - biotinContents

Recombinant Mononucleosomes H3K79me1 (MLA) – biotin (0.5 mg protein + 0.5 mg DNA) are supplied in 10 mM Tris-HCl pH 8.0, 1 mM EDTA, 2 mM DTT, and 20% glycerol.

for Recombinant Mononucleosomes H3K79me1 (MLA) - biotinBackground

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 H3K79me1 (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 cϒ-provides precise control over the site and degree of methylation. The MLA technology is covered under U.S. Patent No. 8,278,112.

for Recombinant Mononucleosomes H3K79me1 (MLA) - biotinApplication Notes

Recombinant Mononucleosomes H3K79me1 (MLA) - biotin 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.

for Recombinant Mononucleosomes H3K79me1 (MLA) - biotinProtein Details

Recombinant Mononucleosomes H3K79me1 (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 monomethyl Lys79 (H3K79me1) (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 H3K79me1 (MLA) - biotin SDS-PAGE gel

Recombinant Mononucleosomes H3K79me1 (MLA) - biotin SDS-PAGE gel
12.5% SDS-PAGE Coomassie staining
MW: 108 kDa
Purity: >95%

Pull down assay for Recombinant Mononucleosomes H3K79me1 (MLA) - biotin

Pull down assay for Recombinant Mono-nucleosomes H3K79me1 (MLA) - biotin
Recombinant Mononucleosomes H3K79me1 (MLA) - biotin were pulled down by streptavidin beads. Input nucleosomes (Lane 2) and the nucleosomes (Lane 3) pulled down by streptavidin were run on a 12.5% SDS-PAGE gel and stained with Coomassie blue. The SDS-PAGE gel result shows that almost all of biotinylated mononucleosomes H3K79me1 (MLA) - biotin are pulled down by streptavidin beads.

Western Blot analysis for Recombinant Mono-nucleosomes H3K79me1 (MLA) - biotin

Western Blot analysis for Recombinant Mono-nucleosomes H3K79me1 (MLA) - biotin
Recombinant Mononucleosomes H3K79me1 (MLA) - biotin (Lane 1) and unmodified nucleosomes (Lane 2) were detected with H3K79me1 antibody and H4 antibody, respectively. H4 was detected as loading control. Only Recombinant Mononucleosomes H3K79me1 (MLA) - biotin can be detected by H3K79me1 antibody.

for Recombinant Mononucleosomes H3K79me1 (MLA) - biotinStorage

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.

for Recombinant Mononucleosomes H3K79me1 (MLA) - biotinGuarantee

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.

 

Technical Data Sheet

Recombinant Mononucleosomes H3K79me1 (MLA) - biotin
 

Data Thumbnails

Recombinant Mononucleosomes H3K79me1 (MLA) - biotin SDS-PAGE gel

Recombinant Mononucleosomes H3K79me1 (MLA) - biotin SDS-PAGE gel
(Click to enlarge and view details)

Pull down assay for Recombinant Mononucleosomes H3K79me1 (MLA) - biotin

Pull down assay for Recombinant Mono-nucleosomes H3K79me1 (MLA) - biotin
(Click to enlarge and view details)

Western Blot analysis for Recombinant Mono-nucleosomes H3K79me1 (MLA) - biotin

Western Blot analysis for Recombinant Mono-nucleosomes H3K79me1 (MLA) - biotin
(Click to enlarge and view details)