Histone Octamer (H3.3), Human Recombinantt Description: Human histone octamers assembled from recombinant human histones expressed in E. coli (two each of histones H2A, H2B, H3.3 and H4; accession numbers: H2A-P04908; H2B-O60814; H3.3-P84243; H4-P62805). The histone octamer is the protein component of the nucleosome, the basic subunit of chromatin. A nucleosome consists of a histone octamer wrapped with 147 base pairs of DNA (note that there is only histone octamer in this product).
Histone Octamer (H3.3), Human Recombinantt Formulation: Gel filtration-purified recombinant human histone octamers ( mg/ml) in μl of 10 mM Tris-HCl pH 7.5, 1 mM EDTA, 2 M NaCl, 2 mM DTT, 20% glycerol. MW: 108,392.2 Da. Molarity: μM.
Histone Octamer (H3.3), Human Recombinantt Storage and Stability: Stable for six months at -80°C from date of receipt. For best results, aliquot and avoid multiple freeze/thaws.
Histone Octamer (H3.3), Human Recombinantt Application Notes: Human recombinant histone octamers can be used for chromatin reconstitution experiments, or as substrates for histone modifying enzymes. See product page on web for links to protocols for chromatin reconstitution. Histone octamers should not be stored under conditions where the salt concentration is lower than 800 mM. Please note that the concentration may vary compared to previous lots.
References: Lowary PT and J Widom (1998). J Mol Biol 276: 19-42. Link Luger K et al (1999). Methods Mol Biol 119: 1-16. Link
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DNA Gel Data: Histone Octamer (H3.3), Human Recombinantt run on an agarose gel and stained with ethidium bromide to visualize DNA. Lane 1: Intact nucleosomes (200 ng total, ~100 ng DNA and ~100 ng histone protein) run on gel. Lane 2: DNA extracted from nucleosomes (400 ng). Intact nucleosomes migrate much higher than free DNA, thus the DNA resolves at a higher molecular weight when nucleosome-bound. Note absence of free DNA in Lane 1, indicating all DNA complexed with histones.
Protein Gel Data: Coomassie stained PAGE gel of proteins in Histone Octamer (H3.3), Human Recombinant (2 μg) to demonstrate the purity of the histones in the preparation. Sizes of molecular weight markers and positions of the core histones (H2A, H2B, H3.3 and H4) are indicated.