Mononucleosomes (H3.3 N32), Recombinant Human Biotinylated
Mononucleosomes (H3.3 ΔN32), Recombinant Human Biotinylated Description: Mononucleosomes 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), with the amino acid sequence of H3 begining with glycine 33 (amino acids 1-32 are deleted), wrapped by 147 base pairs of 601 positioning sequence DNA. 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.
Mononucleosomes (H3.3 ΔN32), Recombinant Human BiotinylatedFormulation: Mononucleosomes (H3.3 ΔN32), Recombinant Human (27.4 µg protein weight, 50 µg DNA+protein) in 10 mM Tris HCl, pH 7.5, 25 mM NaCl, 1 mM EDTA, 2 mM DTT, 20% glycerol. MW = 193,766.7 Da.
Mononucleosomes (H3.3 ΔN32), Recombinant Human Biotinylated Storage and Stability: Stable for six months at -80°C from date of receipt. For best results, aliquot and avoid multiple freeze/thaws.
Mononucleosomes (H3.3 ΔN32), Recombinant Human Biotinylated Application Notes: Mononucleosomes (H3.3 ΔN32), Human Recombinant Biotinylated are highly purified and suitable for use as substrates in enzyme screening assays, structural studies, or effector protein binding experiments. The N-terminal deletion allows for the study of the role of the N-terminus in many aspects of chromatin biology.
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Protein Gel Data: Coomassie stained PAGE gel of proteins in Monoucleosomes (H3.3 ΔN32), Human Recombinant Biotinylated (1 µg) demonstrates the purity of the histones in the preparation. Sizes of molecular weight markers and positions of the core histones (H2A, H2B, H3.3 ΔN32 and H4) are indicated. H3.3 ΔN32 and H4 co-migrate. (Click image to enlarge)
DNA Gel Data: Monoucleosomes (H3.3 ΔN32), Human Recombinant Biotinylated run on an agarose gel and stained with ethidium bromide to visualize DNA. Lane 1: DNA extracted from nucleosomes (100 ng). Lane 2: Intact nucleosomes (300 ng). (Click image to enlarge)
Western Blot Data: Western Analysis of Nucleosome (H3.3 ΔN32), Human Recombinant Biotinylated. Top Panel: Wild type (Lane 1) and H3.3 ΔN32-containing nucleosomes (Lane 2) were probed with an anti-H3 COOH-terminal antibody and analyzed via ECL readout. Bottom Panel: Detail from Coomassie stained gel showing histones from H3.3 WT (Lane 1) and H3.3 ΔN32 nucleosomes (Lane 2). (Click image to enlarge)