Mononucleosomes, Human Recombinant Formulation: Mononucleosomes assembled from recombinant human histones expressed in E. coli (two each of histones H2A, H2B, H3 and H4. Accession numbers: H2A-P04908; H2B-O60814; H3.1-P68431; H4-P62805) wrapped by 147 base pairs of 601 positioning sequence DNA. The nucleosome is the basic subunit of chromatin.
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Mononucleosomes, Human Recombinant Formulation: Purified recombinant mononucleosomes (100 µg total mass, protein+DNA, 54.6 µg protein) in 10 mM Tris-HCl pH 7.5, 1 mM EDTA, 25 mM NaCl, 2 mM DTT, & 20% glycerol.
Mononucleosomes, Human Recombinant Storage and Stability: Stable for six months at -20°C from date of receipt. For best results, aliquot and avoid multiple freeze/thaws.
Mononucleosomes, Human Recombinant Application Notes: Mononucleosomes, Human Recombinant are highly purified and are suitable for use as substrates in enzyme screening assays or for nucleosome binding experiments. The absence of post-translational histone modifications makes them ideal for conducting enzyme activity and screening assays. EpiCypher Mononucleosomes, Human Recombinant do not contain free DNA which could alter assayed activities.
References: Strelow JM et al (2016). J Biomol Screen 21: 786-794. Link Munari F et al (2012). J Biol Chem 287: 33756-33765. Link Lowary PT and J Luger K et al (1999). Methods Mol Biol 119: 1-16. Link Widom (1998). J Mol Biol 276: 19-42. Link
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DNA Gel Data: Mononucleosomes, Recombinant Human run on an agarose gel and stained with ethidium bromide to visualize DNA. Lane 1: Free DNA extracted from nucleosomes (200 ng). Lane 2: Intact nucleosomes (400 ng). Click image to enlarge
Protein Gel Data: Coomassie stained PAGE gel of proteins in Mononucleosomes, Human Recombinant (0.5 µg) to demonstrate the purity and integrity of the histones in the preparation. Sizes of molecular weight markers and positions of the core histones (H2A, H2B, H3 and H4) are indicated. Click image to enlarge
HMTase Assay Data: Mononucleosomes, Recombinant Human used as a substrate in a radioactive histone methyltransferase assay with (+SET8) or without (-SET8) recombinant SET8, which is an HMTase that requires nucleosomal substrates for activity. Click image to enlarge