Nucleosome, Recombinant Human, H3 Cit2,8,17 dNuc 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) wrapped by 147 base pairs of 601 positioning sequence DNA. Histone H3 (created by a proprietary synthetic method) contains citrulline (instead of arginine) at positions 2, 8, and 17. The nucleosome is the basic subunit of chromatin. The 601 sequence, identified by Lowary and Widom, is a 147-base pair sequence that has high affinity for histone octamers and is useful for nucleosome assembly and contains a 5' biotin-TEG group.
Nucleosome, Recombinant Human, H3 Cit2,8,17 dNuc BiotinylatedFormulation: Recombinant H3 Cit2,8,17 mononucleosomes (27.4 µg protein weight, 50 µg DNA+protein) in 10 mM Tris-HCl pH 7.5, 1 mM EDTA, 25 mM NaCl, 2 mM DTT, & 20% glycerol. Nucleosome molecular weight = 200,955 Da.
Nucleosome, Recombinant Human, H3 Cit2,8,17 dNuc 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, H3 Cit2,8,17 dNuc Biotinylated Application Notes: Nucleosome, Recombinant Human, H3 Cit2,8,17 are highly purified and are suitable for use as substrates in enzyme screening assays or for effector protein binding experiments.
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Western Blot Data: Western Analysis of Nucleosome, Recombinant Human, H3Cit2,8,17. Top Panel: Unmodified H3 (Lane 1) and H3Cit2,8,17 containing nucleosomes (Lane 2) were probed with an anti-citrulline antibody and analyzed via ECL readout. Only the H3Cit2,8,17 sample produced a detectable signal. Bottom Panel: Detail from Coomassie stained gel showing unmodifed H3 nucleosome (Lane 1) and H3Cit2,8,17 nucleosome (Lane 2). (Click image to enlarge)
Mass Spec Data: Synthetic H3 Cit2,8,17 protein analyzed by ESI-TOF mass spectrometry. Expected mass = 15,300.9 Da. Determined mass = 15,300.96 Da. (Click image to enlarge)
Protein Gel Data: Coomassie stained PAGE gel of proteins in Nucleosome, Recombinant Human, H3Cit2,8,17 (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, H3Cit2,8,17 and H4) are indicated. (Click image to enlarge)
DNA Gel Data: Nucleosome, Recombinant Human, H3Cit2,8,17 resolved by native PAGE and stained with ethidium bromide to visualize DNA. Lane 1: Free DNA extracted from nucleosomes (100 ng). Lane 2: Intact nucleosomes (250 ng). (Click image to enlarge)