Tailless Nucleosomes, Human Recombinant, Biotinylated Description: 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). All four of the histones have had their tails enzymatically removed after wrapping by 147 base pairs of 601 positioning sequence DNA. There is a 5’ biotin-TEG group on the DNA which makes these nucleosomes ideal for use as a negative control in binding assays and pull-down experiments. The nucleosome is the basic subunit of chromatin.
Tailless Nucleosomes, Human Recombinant, Biotinylated Formulation: Purified recombinant tailless nucleosomes (50 µg total mass protein+DNA, 25 µg protein) in 10 mM Tris-HCl pH 7.5, 1 mM EDTA, 25 mM NaCl, 2 mM DTT, & 20% glycerol.
Tailless Nucleosomes, Human Recombinant, Biotinylated Storage and Stability: Stable for six months at -80°C from date of receipt. For best results, aliquot and avoid multiple freeze/thaws.
Tailless Nucleosomes, Human Recombinant, Biotinylated Application Notes: Tailless Nucleosomes, Human Recombinant, Biotinylated are highly purified and are suitable for use as a negative control or substrate in enzyme screening assays or for nucleosome binding experiments. The absence of histone post-translational modifications makes them ideal for conducting enzyme activity and screening assays. The biotin group on the DNA makes pull-down experiments possible, allowing isolation of the nucleosomes after assay completion. EpiCypher Tailless Nucleosomes, Human Recombinant, Biotinylated do not contain free DNA which could alter assayed activities.
References: Strelow JM et al (2016) J Biomol Screen 21: 786-794. Link
Munari F et al (2012) J Biol Chem 287: 33756-33765. Link
Luger K et al (1999) Methods Mol Biol 119: 1-16. Link
Lowary PT and J Widom (1998) J Mol Biol 276: 19-42. Link
Ausio J et al (1989) J Mol Biol 206(3): 451-63. Link
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Protein Gel Data: Coomassie stained SDS-PAGE to demonstrate purity of the histones in the preparation. Lane 1 shows histone proteins in the nucleosome before enzymatic removal of tails. Lane 2 shows the resulting histone proteins after tail removal. Size of molecular weight markers and positions of the intact core histones (H2A, H2B, H3, and H4) are indicated.
Click image to enlarge.
DNA Gel Data: Recombinant Nucleosomes resolved via native PAGE and stained with ethidium bromide to visualize DNA. Lane 1: Free DNA (100ng). Lane 2: Intact Nucleosomes (400ng). Lane 3: Tailless Nucleosomes after enzymatic removal of histone tails. Click image to enlarge.