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Recombinant Proteins & Enzymes

Recombinant Mononucleosomes H3T3ph (EPL)

Catalog No: 81161 Format: 20 µg $415 Buy Now
Catalog No: 81861 Format: 1 mg $6,225 Buy Now
Expressed In: E. coli Protein Species: Human

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Expressed Protein Ligation

Recombinant Mononucleosomes H3T3ph (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 H3T3ph (EPL)Contents

Recombinant Mononucleosomes H3T3ph (EPL) (20 µg protein + 20 µg DNA) are supplied at a protein concentration of 0.55 μg/μl in 10 mM Tris-HCl pH 8.0, 1 mM EDTA, 2 mM DTT and 20% glycerol.

for Recombinant Mononucleosomes H3T3ph (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 H3T3ph (EPL)Application Notes

Recombinant Mononucleosomes H3T3ph (EPL) are suitable for use in the study of enzyme kinetics, inhibitor screening, and selectivity profiling.

for Recombinant Mononucleosomes H3T3ph (EPL)Protein Details

Recombinant Mononucleosomes H3T3ph (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 phosphorylation at threonine 3, 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. H3T3ph (Histone H3 phosphoryl Thr3) 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 an N-terminal histone tail peptide containing phosphoryl threonine 3 via a native peptide bond.

 
Recombinant Mononucleosomes H3T3ph (EPL) SDS PAGE

Recombinant Mononucleosomes H3T3ph (EPL) SDS PAGE
13% SDS-PAGE gel, stained with Coomassie blue
Purity: ≥ 92%

Recombinant Mononucleosomes H3T3ph (EPL) DNA gel

Recombinant Mononucleosomes H3T3ph (EPL) DNA gel
Mononucleosomes H3T3ph (EPL) were run on a 2% agarose gel and stained with ethidium bromide. Lane 1: DNA marker. Lane 2: Intact mononucleosome H3T3ph. Lane 3: 601 DNA. Intact mononucleosome H3T3ph migrated higher than free 601 DNA. The agarose gel result shows almost all of 601 DNA wraps histone octamers to form nucleosomes.

Western Blot analysis for Recombinant Mononucleosomes H3T3ph (EPL)

Western Blot analysis for Recombinant Mononucleosomes H3T3ph (EPL)
Recombinant Mononucleosomes H3T3ph (EPL) (Lane 1) and unmodified nucleosomes (Lane 2) were detected with H3T3ph antibody and H4 antibody, respectively. H4 was detected as loading control. Only Recombinant Mononucleosomes H3T3ph (EPL) can be detected by H3T3ph antibody.

for Recombinant Mononucleosomes H3T3ph (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 H3T3ph (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 H3T3ph (EPL)
 

Data Thumbnails

Recombinant Mononucleosomes H3T3ph (EPL) SDS PAGE

Recombinant Mononucleosomes H3T3ph (EPL) SDS PAGE
(Click to enlarge and view details)

Recombinant Mononucleosomes H3T3ph (EPL) DNA gel

Recombinant Mononucleosomes H3T3ph (EPL) DNA gel
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Western Blot analysis for Recombinant Mononucleosomes H3T3ph (EPL)

Western Blot analysis for Recombinant Mononucleosomes H3T3ph (EPL)
(Click to enlarge and view details)