Nucleosome, Recombinant Human, H3K27ac 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 acetyl-lysine at position 27. 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, H3K27ac dNuc BiotinylatedFormulation: Recombinant H3K27ac 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 = 200,774 Da.
Nucleosome, Recombinant Human, H3K27ac 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, H3K27ac dNuc Biotinylated Application Notes: Nucleosome, Recombinant Human, H3K27ac are highly purified and are suitable for use as substrates in enzyme screening assays or for effector protein (bromo and YEATS domains) binding experiments.
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Western Blot Data: Western Analysis of Nucleosome, Recombinant Human, H3K27ac. Top Panel: Unmodified H3 (Lane 1) and H3K27ac containing nucleosomes (Lane 2) were probed with an anti-H3K27ac antibody and analyzed via ECL readout. Only the H3K27ac sample produced a detectable signal. Bottom Panel: Detail from Coomassie stained gel showing unmodified nucleosomes (Lane 1) and H3K27ac nucleosomes (Lane 2). (Click image to enlarge)
Mass Spec Data: Synthetic H3K27ac protein analyzed by ESI-TOF mass spectrometry. Expected mass = 15,314.9 Da. Determined mass = 15,315.0 Da. (Click image to enlarge)
Protein Gel Data: Coomassie stained SDS-PAGE gel of proteins in Nucleosome, Recombinant Human, H3K27ac (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, H3K27ac and H4 are indicated. (Click image to enlarge)
DNA Gel Data: Nucleosome, Recombinant Human, H3K27ac examined by native PAGE and stained with ethidium bromide to visualize DNA. Lane 1: Free DNA (200 ng). Lane 2: Intact Nucleosome, Recombinant Human, H3K27ac (400 ng). (Click image to enlarge)