Recombinant Mononucleosomes (H3.3) - biotinylated (20 μg protein + 20 μg DNA) is 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.
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.
Recombinant Mononucleosomes (H3.3) - biotinylated are suitable for use in the study of enzyme kinetics, inhibitor screening, and selectivity profiling.
HMT Assay Conditions: 2 μg mononucleosomes was incubated with different concentration of NSD2-SET (Cat# 31476) in reaction buffer including 50 mM TrisCl, pH 8.6, 0.02% Triton X-100, 2 mM MgCl2, 1 mM TCEP, 50 μM SAM for 3 hours at room temperature. Activity was detected by Western blot.
Recombinant Mononucleosomes (H3.3) - biotinylated consists of 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 NM_003512), H2B that includes amino acids 1-126 (end) (accession number NM_003518), H3.3 that includes amino acids 1-136 (end) (accession number NM_005324), 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. The recombinant protein is >95% pure by SDS-PAGE.
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.
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.