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Recombinant Proteins & Enzymes

Recombinant Mononucleosomes H3K9ac (EPL)

Catalog No: 81075 Format: 20 µg $415 Buy Now
Expressed In: E. coli Protein Species: Human

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Expressed Protein Ligation

Recombinant Mononucleosomes H3K9ac (EPL) has been generated using the patented Expressed Protein Ligation (EPL) technology. EPL can be used to generate methylated, acetylated or phosphorylated histones. In EPL, the histone globular domain is ligated to a peptide that contains the N-terminal histone tail with the desired site-specific modification. The ligation reaction maintains the native histone bonds, and the resulting recombinant protein more closely mimics natural histones.

for Recombinant Mononucleosomes H3K9ac (EPL)Contents

Recombinant Mononucleosomes H3K9ac (EPL) (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. 

for Recombinant Mononucleosomes H3K9ac (EPL)Background

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 H3K9ac (EPL)Application Notes

Recombinant Mononucleosomes H3K9ac (EPL) are suitable for use in the study of enzyme kinetics, inhibitor screening, and selectivity profiling.

for Recombinant Mononucleosomes H3K9ac (EPL)Protein Details

Recombinant Mononucleosomes H3K9ac (EPL) consist of a 167 bp of 601 DNA 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), H3.2 that includes amino acids 1-136 (end) (accession number NP_066403.2) with acetylation at lysine 9, and H4 that includes amino acids 1-103 (end) (accession number NP_003539.1). All of these histones were expressed in E. coli cells. The molecular weight of histone octamer is ~108 kDa.

H3K9ac (Histone H3 acetyl Lys9) protein is generated using expressed protein ligation (EPL) technology. Truncated human Histone H3.2 is produced in E. coli and purified using FPLC. The purified protein is subsequently ligated to a N-terminal histone tail peptide containing acetyl lysine 9 via a native peptide bond.

 
Recombinant Mononucleosomes H3K9ac (EPL) SDS PAGE

Recombinant Mononucleosomes H3K9ac (EPL) SDS PAGE
13% SDS-PAGE Coomassie staining
Purity: ≥ 95% 

Recombinant Mononucleosomes H3K9ac (EPL) DNA gel

Recombinant Mononucleosomes H3K9ac (EPL) DNA gel
Mononucleosomes H3K9ac (EPL) were run on a 2% agarose gel and stained with ethidium bromide. Lane 1: DNA marker. Lane 2: 601 DNA. Lane 3: Intact nucleosome. Intact nucleosomes migrate much higher than free 601 DNA. The agarose gel result shows almost all of 601 DNA wraps histone octamers to form nucleosomes. 

Recombinant Mononucleosomes H3K9ac (EPL) tested by Western blot.

Western Blot analysis for Recombinant Mono-nucleosomes H3K9ac (EPL)
Unmodified nucleosomes (Lane 1) and Recombinant Mononucleosomes H3K9ac (EPL) (Lane 2) were detected with H3K9ac antibody (Cat. No. 39137) and H4 antibody (Cat. No. 61300), respectively. H3 was detected as loading control. Only Recombinant Mononucleosomes H3K9ac (EPL) can be detected by anti-H3K9ac antibody.

for Recombinant Mononucleosomes H3K9ac (EPL)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.

for Recombinant Mononucleosomes H3K9ac (EPL)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.

 

Technical Data Sheet

Recombinant Mononucleosomes H3K9ac (EPL)
 

Data Thumbnails

Recombinant Mononucleosomes H3K9ac (EPL) SDS PAGE

Recombinant Mononucleosomes H3K9ac (EPL) SDS PAGE
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Recombinant Mononucleosomes H3K9ac (EPL) DNA gel

Recombinant Mononucleosomes H3K9ac (EPL) DNA gel
(Click to enlarge and view details)

Recombinant Mononucleosomes H3K9ac (EPL) tested by Western blot.

Western blot of Recombinant Mononucleosomes H3K9ac (EPL).
(Click image to enlarge and see details.)