Nucleosome, Recombinant Human, H3K14ub1 dNuc, Biotinylated

SKU: 16-0398
Pack Size: 25 µg
  • Species:  Human
  • Source:  E. coli & synthetic DNA
  • Tag:  Biotinylated
  • Molecular Weight:  216,857.3 Da


H3K14ub1 designer mononucleosomes (dNucs) consist of 147 base pairs of DNA wrapped around an octamer core of histone proteins (two each of H2A, H2B, H3.1 and H4) to form a nucleosome, the basic repeating unit of chromatin. H3K14ub1 has been shown to promote the activity of the methyltransferase Clr4, which methylates H3K9, a known repressive mark [1]. The 147 bp 601 sequence, identified by Lowary & Widom [2], has high affinity for histone octamers and is useful for nucleosome assembly. The DNA in this nucleosome contains a 5’ biotin-TEG group. H3K14ub1 has a Cys to Ala substitution at position 110.

Validation Data

Figure 1: DNA gel data
H3K14ub1 dNuc resolved via native PAGE gel and stained with ethidium bromide to visualize DNA. Lane 1: Free DNA (EpiCypher 18-0005; 100 ng). Lane 2: Intact H3K14ub1 nucleosomes (400 ng). NOTE: The split band for the H3K14ub1 is a gel running artifact of the ubiquitin impacting the resolution of the nucleosome.

Figure 2: Mass spec data
Semi-synthetic H3K14ub1 histone analyzed by high resolution mass spectrometry. Expected mass = 23,771.60 Da. Determined mass = 23,772.47 Da.

Figure 3: Protein gel data
Coomassie stained SDS-PAGE gel of proteins in H3K14ub1 dNuc (1 µg) demonstrates the purity of histones in the preparation. Sizes of molecular weight markers and positions of the core histones (H2A, H2B, H3K14ub1 and H4) are indicated.

Technical Information

Stable for six months at -80°C from date of receipt. For best results, aliquot and avoid freeze/thaws.
10 mM Tris HCl pH 7.5, 25 mM NaCl, 1 mM EDTA, 2 mM DTT, 20% glycerol. (14.4 µg protein, 25 µg DNA + protein).

Application Notes

H3K14ub1 dNuc is highly purified and suitable for a variety of applications, including use as a substrate in enzyme assays, high-throughput screening and inhibitor testing, chromatin binding studies, protein-protein interaction assays, structural studies, and in effector protein binding experiments.

Gene & Protein Information

UniProt ID

H2A - P04908 (alt. names: H2A type 1-B/E, H2A.2, H2A/a, H2A/m)

H2B - O60814 (alt. names: H2B K, HIRA-interacting protein 1)

H3.1 - P68431 (alt. names: H3, H3/a, H3/b, H3/c, H3/d)

H4 - P62805


Background References:
[1] Oya et al. EMBO Rep. (2019). PMID: 31468675
[2] Lowary & Widom J. Mol. Biol. (1998). PMID: 9514715

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