Nucleosome,

Nucleosome, Recombinant Human, H3K27acS28phos dNuc, Biotinylated

$575.00
SKU: 16-0385
Pack Size: 50 μg
  • Species:  Human
  • Source:  E. coli & synthetic DNA
  • Tag:  Biotinylated
  • MW:  200,103.26 Da

Description

Recombinant mononucleosomes (H3K27acS28phos) 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. The 147 bp 601 sequence, identified by Lowary and Widom [1], has high affinity for histone octamers and is useful for nucleosome assembly. H3K27acS28phos dNuc contains acetyl-lysine at position 27 and phosphoserine at position 28 on histone H3.1. The DNA in this nucleosome contains a 5’ biotin-TEG group.

Validation Data

Figure 1: Protein gel data
Coomassie stained SDS-PAGE gel of proteins in H3K27acS28phos (1 µg) demonstrates the purity of histones in the preparation. Sizes of molecular weight markers and positions of the core histones (H2A, H2B, H3K27acS28phos and H4) are indicated.

Figure 2: Mass spec data
Synthetic H3K27acS28phos histone analyzed by high resolution mass spectrometry. Expected mass = 15,394.9 Da. Determined mass = 15,395.87 Da.

Figure 3: DNA gel data
H3K27acS28phos 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 H3K27acS28phos 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

H3K27acS28phos mononucleosome 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

References

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

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