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Recombinant Proteins & Enzymes

Recombinant Mononucleosomes H3K4me3/K27me3 - biotin

Catalog No: 81005 Format: 10 µg $395 Buy Now
Expressed In: Synthetic Protein Species: Human

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for Recombinant Mononucleosomes H3K4me3/K27me3 - biotinContents

Recombinant Mononucleosomes H3K4me3/K27me3 - biotinylated (10 µg protein + 10 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 H3K4me3/K27me3 - biotinBackground

In vivo, histones are wrapped around by DNA in chromatin. Therefore, nucleosomes are more physiologically relevant substrates than histones and histone-derived peptides for 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.

for Recombinant Mononucleosomes H3K4me3/K27me3 - biotinApplication Notes

Recombinant Mononucleosomes H3K4me3/K27me3)- biotinylated are suitable for use as substrate of enzymatic assay or other biochemical assay.

for Recombinant Mononucleosomes H3K4me3/K27me3 - biotinProtein Details

Recombinant Mononucleosomes H3K4me3/K27me3 - biotinylated consist of a 167 bp of 601 DNA with 5’ biotin tag and two molecules each of histones H2A that includes amino acids 1-130 (end) (accession number NP_003503.1), H2B that includes amino acids 1-126 (end) (accession number NP_003509.1), H3.1 that includes amino acids 1-136 (end) (accession number NP_003520.1) with trimethylation at lysine 4 and lysine 27, and H4 that includes amino acids 1-103 (end) (accession number NP_003539.1). Histone H2A, H2B and H4 were expressed in E. coli cells. Histone H3K4me3/K27me3 was synthesized completely. H3K4me3/K27me3 corresponds to the native histone sequence and does not contain any amino acid substitutions or residue analogs. The molecular weight of histone octamer is ~108 kDa.

 
Mass Spec analysis of Recombinant Mononucleosomes H3K4me3/H3K27me3 - biotin.

Mass Spec analysis of Recombinant Mononucleosomes H3K4me3/H3K27me3 - biotin.
Synthetic H3K4me3/K27me3 protein was analyzed by ESI-TOF mass spectrometry.
Expected mass = 15357.8 Da.
Determined mass = 15357.5 Da.

Recombinant Mononucleosomes H3K4me3/H3K27me3 - biotin, DNA gel.

Recombinant Mononucleosomes H3K4me3/H3K27me3 - biotin, DNA gel.
Recombinant Mononucleosomes H3K4me3/K27me3 - biotin were run on a 2% agarose gel and stained with ethidium bromide. Lane 1: DNA marker. Lane 2: 601 DNA. Lane 3: Intact mononucleosomes H3K4me3/K27me3. Intact mononucleosomes H3K4me3/K27me3 migrated much higher than free 601 DNA. The agarose gel result shows almost all of 601 DNA wrap histone octamers to form nucleosomes.

Western Blot analysis of Recombinant Mononucleosomes H3K4me3/H3K27me3 - biotin.

Western Blot analysis of Recombinant Mononucleosomes H3K4me3/H3K27me3 - biotin.
Unmodified nucleosomes (Lane 1) and Recombinant Mononucleosomes H3K4me3/K27me3 - biotin (Lane 2) were detected with antibody anti-H3K4me3, anti-H3K27me3 and anti-H4, respectively. H4 was detected as loading control. Only Recombinant Mononucleosomes H3K4me3/K27me3 - biotin can be detected by anti-H3K4me3 and anti- H3K27me3 antibody.

for Recombinant Mononucleosomes H3K4me3/K27me3 - 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 H3K4me3/K27me3 - 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 H3K4me3/K27me3 - biotin
 

Data Thumbnails

Mass Spec analysis of Recombinant Mononucleosomes H3K4me3/H3K27me3 - biotin.

Mass Spec analysis of Recombinant Mononucleosomes H3K4me3/H3K27me3 - biotin.
(Click to enlarge and view details).

Recombinant Mononucleosomes H3K4me3/H3K27me3 - biotin, DNA gel.

Recombinant Mononucleosomes H3K4me3/H3K27me3 - biotin, DNA gel.
(Click to enlarge and view details).

Western Blot analysis of Recombinant Mononucleosomes H3K4me3/H3K27me3 - biotin.

Western Blot analysis of Recombinant Mononucleosomes H3K4me3/H3K27me3 - biotin.
(Click to enlarge and view details).