Histones, Control (+/-sodium butyrate)

Applications: \nImmunoblot Analysis: 10ug of histones from untreated and sodium butyrate-treated HeLa cells were used as a positive control for immunoblot analysis using 2ug/ml anti-acetyl Histone H4, 1ug/ml anti-acetyl-Histone H3 and 1:3000 dilution of anti-hyperacetylated Histone H4 (Penta). 10ug Control Histones, untreated or sodium butyrate-treated, were resolved by electrophoresis, transferred to nitrocellulose and probed with 2ug/ml anti-acetyl-Histone H4. Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.\n\nKit Components:\nH5110-06B1 Control Histones, untreated, (HeLa cell acid extract), 10x50ug of precipitated core histones, lyophilized from sterile water. \n\nH5110-06B2 Control Histones, sodium butyrate-treated, (HeLa cell acid extract), 10x50ug of precipitated core histones, lyophilized from sterile water.\n\n\nStorage and Stability: Stable for 2 years at -20 degrees C from date of shipment, when stored with provided desiccant. Stable for 6 months at -70 degrees C when reconstituted. To rehydrate, aseptically reconstitute to 1ug/ml with sterile water. Aliquot to avoid repeated freezing and thawing.\n\n\nAutoantibodies to histone antigens have been described in patients with idiopathic and drug-induced SLE, rheumatoid arthritis, and other conditions. The presence of autoantibodies to histones are frequently found in several rheumatic disorders (1). In one study, the predominant responses to histones in SLE sera were to H1, H2b, and H3. Marked elevations of binding occurred to H1 and H2b in 33% of patients, while 25% showed higher binding to H3 (2). The same study showed the highest anti-histone reactivity to be in patients with rheumatoid arthritis with vasculitis, while the highest reactivity in SLE sera was in those patients with a history of photosensitivity (3).\n\nIn diploid eukaryotic cells, the chromatin fibers are about 20nM in diameter. They consist of two major components in equal amounts, DNA and basic proteins called histones. The histones are a group of water and dilute acid soluble basic proteins found associated with DNA in chromosomes. They are characterized by relatively high levels of lysine and arginine. Although histones are classified into a limited number of types of fractions (see below) with each particular fraction having a fundamentally distinct amino acid composition and sequence, numerous subfractions are observed due to the acetylation, methylation, and phosphorylation of various amino acid residues. Microheterogeneity or alteration of structure is dynamic such that the histones of a single cell type are found to vary during development. They are believed to play a role in gene activity and cellular metabolism.\n\nHistones are believed to be regularly arranged in the deep groove of the DNA helix. The recurring positive charges of the histones form electrostatic associations with the negatively charged phosphate groups of DNA making the DNA more stable and flexible. This allows for the supercoiling of the chromatin fibers.\n\nWith the exception of H1, the primary structures of the calf thymus histones have been determined. Comparisons with the structures for histones from other sources indicate that the histones rank among the most highly conserved (low mutation rate) proteins in nature. \n\nMolecular Weights of Histones:\nLysine Rich (H1, f1): ~ 21,500 \nSlightly Lysine Rich (H2a, f2a2): 14,004 \nSlightly Lysine Rich (H2b, f2b): 13,774 \nArginine Rich (H3, f3): 15,324 \nArginine Rich (H4, f2a1): 11,282

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