Nucleosome,

Nucleosome, Recombinant Human, H3.3K36me3dNuc, Biotinylated

$575.00
SKU: 16-0390
Pack Size: 50 µg
  • Species:  Human
  • Source:  E. coli & synthetic DNA
  • Tag:  Biotinylated
  • Molecular Weight:  199,749.4 Da

Description

EpiCypher dNucs™ are semi-synthetic recombinant nucleosomes, containing one or more histone post translational modifications (PTMs). Nucleosomes, the basic repeating unit of chromatin, are the natural targets of chromatin readers and modifying enzymes, providing a physiologically relevant substrate for characterization of epigenetic proteins. dNucs consist of 147 base pairs of DNA wrapped around an octamer core of histone proteins (two each of H2A, H2B, H3.3, and H4). The 147 bp 601 sequence, identified by Lowary and Widom [1], has high affinity for histone octamers and is useful for nucleosome assembly. H3.3K36me3 dNuc contains trimethylated lysine at position 36 on histone H3.3. The DNA in this nucleosome contains a 5’ biotin-TEG group.

Validation Data

Figure 1: Western blot data
Western Analysis of H3.3K36me3 dNuc. Top Panel: Unmodified (EpiCypher 16-0006; Lane 1) and H3.3K36me3 nucleosomes (Lane 2) were probed with an anti-H3.3K36me3 antibody and analyzed via ECL readout. Only the H3.3K36me3 sample produced a detectable signal. Bottom Panel: Detail from Coomassie stained gel showing unmodified (Lane 1) and H3.3K36me3 nucleosomes (Lane 2).

Figure 2: Mass spec data
Semi-synthetic H3.3K36me3 histone analyzed by high resolution mass spectrometry. Expected mass = 15,238.72 Da. Determined mass = 15,237.99 Da.

Figure 3: Protein gel data
Coomassie stained SDS-PAGE gel of proteins in H3.3K36me3 dNuc (1 µg) demonstrates the purity of histones in the preparation. Sizes of molecular weight markers and positions of the core histones (H2A, H2B, H3.3K36me3 and H4) are indicated.

Figure 4: DNA gel data
H3.3K36me3 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 H3.3K36me3 nucleosomes (400 ng).

Technical Information

Storage
Stable for six months at -80°C from date of receipt. For best results, aliquot and avoid freeze/thaws.
Formulation
10 mM Tris HCl pH 7.5, 25 mM NaCl, 1 mM EDTA, 2 mM DTT, 20% glycerol (27.4 µg protein, 50 µg DNA + protein)

Application Notes

H3.3K36me3 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)

H3.3 - P84243

H4 - P62805

References

Background References:
[1] Lowary & Widom J. Mol. Biol. (1998). PMID: 9514715

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