Nucleosome, Recombinant Human, H2AK119ub, 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; H2BO60814; H3.1-P68431; H4-P62805) wrapped by 147 base pairs of 601 positioning sequence DNA. Histone H2A (created by a proprietary synthetic method) contains ubiquitin-lysine at position 119. The nucleosome is the basic subunit of chromatin. The 601 sequence, identified by Lowary and Widom, is a 147-base pair sequece that has
high affinity for histone octamers and is useful for nucleosome assembly and contains a 5’ biotin-TEG group.
Nucleosome, Recombinant Human, H2AK119ub dNuc Biotinylated Formulation: Nucleosome, Recombinant Human, H2AK119ub (27.3 μg protein weight, 50 μg total weight) in μl of 10 mM Tris
HCl, pH 7.5, 25 mM NaCl, 1 mM EDTA, 2 mM DTT, 20% glycerol. Molarity = μmolar. MW = Da.
Nucleosome, Recombinant Human, H2AK119ub, Biotinylated Storage and Stability: Stable for six months at -80°C from date of receipt. For
best results, aliquot and avoid multiple freeze/thaws.
Nucleosome, Recombinant Human, H2AK119ub, Biotinylated Application Notes: Nucleosome, Recombinant Human, H2AK119ub dNucs are highly purified and are suitable for use as substrates in enzyme screening assays or for effector protein binding experiments. Nucleosome, Recombinant Human, H2AK119ub dNucs from EpiCypher does not
contain free DNA which could alter assayed activities.
References: Lowary PT and J Widom (1998). J Mol Biol 276: 19-42.
Luger K et al (1999). Methods Mol Biol 119: 1-16.
View technical datasheet for this product.
Western Blot Data:Western Analysis of Nucleosome,
Recombinant Human, H2AK119ub. Top Panel: Unmodified H2A (Lane 1) and H2AK119ub containing nucleosomes (Lane 2) were probed with an anti-H2AK119ub antibody and analyzed via ECL readout. Only the H2AK119ub sample produced a detectable signal. Bottom Panel: Detailfrom Coomassie stained gel showing unmodified nucleosomes (Lane 1) and H2AK119ub nucleosomes (Lane 2). (Click image to enlarge)
Mass Spec Data:H2AK119ub protein analyzed by high
resolution mass spectrometry. Expected mass = Da. Determined mass = Da. (Click image to enlarge)
Protein Gel Data: Coomassie stained PAGE gel of proteins in
Nucleosome, Recombinant Human, H2AK119ub (1 μg) to demonstrate the purity of the histones in the preparation. Sizes of molecular weight markers and positions of the core histones (H2AK119ub, H2B, H3.1 and H4) are indicated. (Click image to enlarge)
DNA Gel Data: Nucleosome, Recombinant Human, H2AK119ub run on a native PAGE gel and stained with ethidium bromide to visualize DNA. Lane 1: Free DNA extracted from nucleosomes (100 ng). Lane 2: Intact nucleosomes (400 ng). (Click image to enlarge)