Mononucleosomes,

Mononucleosomes, Recombinant Human

$425.00
SKU: 16-0009
Pack Size: 100 μg

Description

Mononucleosomes, Recombinant Human 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 sequence DNA. The nucleosome is the basic subunit of chromatin. The 601 sequence, identified by Lowary and Widom, has high affinity for histone octamers and is useful for nucleosome assembly.

Validation Data

Figure 1: 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).

Figure 2: 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.

Figure 3: 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.

Technical Information

Formulation:

Purified recombinant mononucleosomes (54.6 µg protein by mass, 100 µg protein+DNA in 39.0 µl) in 10 mM Tris-HCl pH 7.5, 1 mM EDTA, 25 mM NaCl, 2 mM DTT, 20% glycerol. Concentration of nucleosomes is 12.8 µM. Nucleosome molecular weight = 200,253 Da.

Storage and Stability:
Stable for six months at -80°C from date of receipt. For best results, aliquot and avoid multiple freeze/thaws.

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, Recombinant do not contain free DNA which could alter assayed activities.

References

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

Product References:
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