Active Motif,
Tools to analyze nuclear function,
Your CartYour Cart 0 items
    
Recombinant Proteins & Enzymes

Recombinant Mononucleosomes H4K20me3 (MLA) - biotin

Catalog No: 81081 Format: 20 µg $395 Buy Now
Catalog No: 81781 Format: 1 mg $5,200 Buy Now
Expressed In: E. coli Protein Species: Human

Request a quote for a bulk orderBulk Order Button


Methylated Lysine Analog

Recombinant Mononucleosomes H4K20me3 (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 H4K20me3 (MLA) - biotinContents

Recombinant Mononucleosomes H4K20me3 (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. Please refer to product insert upon arrival for lot-specific concentration.

for Recombinant Mononucleosomes H4K20me3 (MLA) - biotinBackground

Histone H4 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 histone, present at the interface between the nucleosome core and DNA entry/exit points.

Recombinant Histone H4K20me3 (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 γ-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 H4K20me3 (MLA) - biotinApplication 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

for Recombinant Mononucleosomes H4K20me3 (MLA) - biotinProtein Details

Recombinant Mononucleosomes H4K20me3 (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 that includes amino acids 1-136 (end) (accession number NP_003520.1), and H4 trimethyl Lys 20 (H4K20me3) (MLA) 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 H4K20me3 (MLA) - biotin pull-down by streptavidin

Recombinant Mononucleosomes H4K20me3 (MLA) - biotin pull-down by streptavidin
Input nucleosomes (Lane 2) and the nucleosomes pulled down by streptavidin (Lane 3) were run on a 13% SDS-PAGE gel and stained with Commassie blue. The SDS-PAGE gel shows that most of biotinylated mononucleosomes H4K20me3 are pulled down by streptavidin beads. * indicates streptavidin. 

Recombinant Mononucleosomes H4K20me3 (MLA) - biotin agarose gel

Recombinant Mononucleosomes H4K20me3 (MLA) - biotin agarose gel
Recombinant Mononucleosomes H4K20me3 (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 H4K20me3. Intact mononucleosomes H4K20me3 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 Mononucleosomes H4K20me3 (MLA) – biotin

Western Blot analysis for Recombinant Mononucleosomes H4K20me3 (MLA) – biotin
Unmodified nucleosomes (Lane 1) and Recombinant Mononucleosomes H4K20me3 (MLA) - biotin (Lane 2) were detected with anti-H4K20me3 antibody (Cat. No. 39671) and anti-H3 antibody (Cat. No. 61475), respectively. H3 was detected as loading control. Only Recombinant Mononucleosomes H4K20me3 (MLA) - biotin can be detected by H4K20me3 antibody.

for Recombinant Mononucleosomes H4K20me3 (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 H4K20me3 (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 H4K20me3 (MLA) - biotin
 

Data Thumbnails

Recombinant Mononucleosomes H4K20me3 (MLA) - biotin pull-down by streptavidin

Recombinant Mononucleosomes H4K20me3 (MLA) - biotin pull-down by streptavidin
(Click to enlarge and view details).

Recombinant Mononucleosomes H4K20me3 (MLA) - biotin agarose gel

Recombinant Mononucleosomes H4K20me3 (MLA) - biotin agarose gel
(Click to enlarge and view details).

Western Blot analysis for Recombinant Mononucleosomes H4K20me3 (MLA) – biotin

Western Blot analysis for Recombinant Mononucleosomes H4K20me3 (MLA) – biotin
(Click to enlarge and view details).