Mononucleosomes, Recombinant Human Biotinylated

SKU: 16-0006
Pack Size: 50 μg


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. There is a 5’ biotin-TEG group on the DNA which makes them ideal for use in nucleosome binding assays and pull-down experiments. The nucleosome is the basic subunit of chromatin.

Validation Data

Figure 1: DNA Gel Data
Mononucleosomes, Human Recombinant Biotinylated run on an agarose gel and stained with ethidium bromide to visualize DNA. Lane 1: Intact nucleosomes (400 ng). Lane 2: DNA extracted from nucleosomes (100 ng).

Figure 2: Protein Gel Data
Coomassie stained PAGE gel of proteins in Mononucleosomes, Human Recombinant Biotinylated (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.

Figure 3: Histone Methyltransferase Assay Data
Recombinant SET8 histone methyltransferase was used in radioactive methyltransferase assays with a variety of substrates as indicated, with SET8 exhibiting the highest level of activity in combination with recombinant nucleosomes (rNucs).

Technical Information

Purified recombinant mononucleosomes (50 µg total mass protein+DNA, 27.3 µg protein) in 10 mM Tris-HCl pH 7.5, 1 mM EDTA, 25 mM NaCl, 2 mM DTT, & 20% glycerol.
Storage and Stability
Stable for six months at -20°C from date of receipt. For best results, aliquot and avoid multiple freeze/thaws.

Application Notes

Application Notes Addendum: Mononucleosomes, Human Recombinant Biotinylated 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. The biotin group on the DNA makes pull-down experiments possible, allowing you to isolate the nucleosomes after your assay is complete. EpiCypher Mononucleosomes, Recombinant Human Biotinylated do not contain free DNA which could alter assayed activities.


Background References:
Luger et al. Methods Mol. Biol. (1999) PMID: 10804500
Lowary & Widom J Mol. Biol. (1998) PMID: 9514715

Product References:

Documents & Resources

Current stock: 0