Nucleosome, Recombinant Human, H3.1K27me3,S28phos, Biotinylated

SKU: 16-0397
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 containing a 5’ biotin-TEG. The 601 sequence, identified by Lowary and Widom [1], has high affinity for histone octamers and is useful for nucleosome assembly. Histone H3 (created by a proprietary synthetic method) contains trimethyl-lysine at position 27 and phosphoserine at position 28. H3S28 is an essential residue for modulating PRC2-mediated methylation at H3K27me3 [2].

Validation Data

Figure 1: Protein Gel Data
Coomassie stained PAGE gel of proteins in Nucleosome, Recombinant Human, H3.1K27me3,S28phos, Biotinylated (1 µg) to demonstrate purity of histones in the preparation. Sizes of molecular weight markers and positions of the core histones (H2A, H2B, H3K27me3,S28phos and H4) are indicated.

Figure 2: DNA Gel Data
Nucleosome, Recombinant Human, H3K27me3,S28phos dNucs resolved via native PAGE and stained with ethidium bromide to visualize DNA. Lane 1: Free DNA (100 ng). Lane 2: Intact H3K27me3,S28phos containing nucleosome (400 ng).

Figure 3: Mass Spec Data
Synthetic H3.1K27me3,S28phos protein analyzed by high resolution mass spectrometry. Expected mass = 15,394.9 Da. Determined mass = 15,394.48 Da.

Technical Information

Nucleosome, Recombinant Human, H3.1K27me3,S28phos, Biotinylated in 10 mM Tris HCl pH 7.5, 25 mM NaCl, 1 mM EDTA, 2 mM DTT, 20% glycerol. MW = 200,103.9 Da.
Storage and Stability
Stable for six (6) months at -80°C from date of receipt. For best results, aliquot and avoid multiple freeze/thaws.

Application Notes

H3.1K27me3,S28phos dNuc is highly purified and suitable for a variety of applications, including use as a substrate in enzymatic assays or for effector protein binding experiments.


Background References:
[1] Lowary & Widom J. Mol. Biol. (1998). PMID: 9514715
[2] Yung et al. Cell Rep. (2015). PMID: 26004180

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