| 货号 | 31466-AM |
| 使用方法 | Recombinant Nucleosomes (H3.1) is suitable for use in the study of enzyme kinetics, inhibitor screening, and selectivity profiling. HMT Assay Conditions: 50 mM TrisCl, pH 8.6, 0.02% Triton X-100, 2 mM MgCl2, 1 mM TCEP, 100 μM SAM, 30 ng/μl Recombinant Nucleosomes (H3.1), 30 ng/μl DOT1L (1-416 aa) at 2 hours at room temperature. Activity was detected by fluorography. |
| 供应商 | Active Motif |
| 背景 | 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. |
| 运输条件 | Dry Ice |
| 存放说明 | 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. |
| 外观 | Recombinant Nucleosomes (H3.1) (20 µg protein + 24 µg DNA) is supplied at a concentration of 0.55 µg/µl in 10 mM Tris-HCl, pH 8.0, 1 mM EDTA, 2 mM DTT and 20% glycerol. |
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Recombinant Nucleosomes (H3.1) protein gel. Recombinant Nucleosomes (H3.1) run on an SDS-PAGE gel and stained with Coomassie blue. | |
Recombinant Nucleosomes (H3.1) protein DNA Gel-shift assay Nucleosome (H3.1) and free plasmid DNA were run on a 1% agarose gel and stained with ethidium bromide. Intact oligonucleosomes migrate higher than free DNA, thus the DNA resolves at a higher molecular weight when nucleosome-bound. * represents the mixture of DNA and histone octamers (incorrect assembly). | |
Recombinant Nucleosomes (H3.1) protein activity assay. Results show that nucleosomes are better substrates than octamers for NSD1-SET and NSD2-SET. Lanes 5-9 represent nucleosomes that have been subjected to subsequent freeze-thaw cycles to demonstrate their stability. Activity was detected by fluorography. |
