Ptmbiolabs/Acetyl-Histone H4 (Lys16) rabbit pAb-ChIP Grade/100 μl/GC-132

Product Description

Size: 100 μlAb Type: PolyclonalReactivity: EukaryotesImmunogen: Acetylated H4 (Lys16)-KLHHost: Rabbit

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Product DescriptionThis product is produced by immunizing rabbits with a synthetic acetyl peptide corresponding to residues surrounding Lys16 of human histone H4. Antibodies are purified by protein A-conjugated agarose followed by acetylated histone H4 (Lys16) peptide affinity chromatography.SpecificityThis antibody detects histone H4 only when it is acetylated at Lys16. This antibody has been shown to selectively recognize acetylated H4 peptide at Lys16, but not the acetylated H4 peptide at Lys8 or Lys12, propionylated peptide at Lys12 or Lys16, butyrylated peptide at Lys8, Lys12 or Lys16, crotonylated peptide at Lys8 or Lys16 and the unmodified H4 peptide at Lys16.Recommended ApplicationsChIP, IP. Not tested in other applications*IP=Immunoprecipitation ChIP=Chromatin precipitationRecommended antibody dilutionIP: 1:25 ChIP: 1:25 (4 μl of antibody for 1x 105 HeLa)Important NotesUse at an assay dependent concentration. Optimal dilutions/concentrations should be determined by the end user.

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 A. Dot blotting analysis on indicated amount of acetylated H4 peptide at Lys16 (lane 1), propionylated H4 peptide at Lys16 (lane2), butyrylated H4 peptide at Lys16 (lane 3), crotonylated peptide at Lys16 (lane 4), acetylated H4 peptide at Lys12 (lane 5), propionylated H4 peptide at Lys12 (lane 6), butyrylated H4 peptide at Lys12 (lane 7), acetylated H4 peptide at Lys8 (lane 8), butyrylated H4 peptide at Lys8 (lane 9), crotonylated peptide at Lys8 (lane 10) and unmodified H4 peptide at Lys16 (lane 11) using acetyl-histone H4 (Lys16) rabbit pAb.B. Sonicated chromatin prepared from 1x 105 HeLa cells treated with sodium butyrate was subjected to chromatin immunoprecipitation (ChIP) using either 4µl acetyl-histone H4 (Lys16) rabbit pAb or 1 μg of normal rabbit IgG as a negative control. Real time quantitative PCR was performed on immunoprecipitated DNA using primers specific for the human GAPDH Coding D2 and α Satellite. Data are presented as enrichment of each IPed sample relative to total amount of input chromatin at each amplicon.

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Scientific DescriptionThe ε-amino lysine acetylation of proteins is an important reversible modification controlling protein activity. The amino-terminal tails of core histones undergo lysine acetylation in multiple sites, termed as “histone code”. Lysine acetylation in core histones occurs in response to various stimuli and plays vital roles in the regulation of many cellular processes including chromatin dynamics, DNA transcription, cell cycle progression, apoptosis, differentiation and nuclear import et al. In most species, histone H2A is primarily acetylated at Lys16, 9, 15 and 36; H2B is primarily acetylated at Lys16, 12, 15, 16 and 20. Histone H3 is primarily acetylated at Lys4, 9, 14, 18, 23, 27, 56 and 79. Histone H4 is primarily acetylated at Lys5, 8, 12, 16 and 20. More than 20 histone acetyltransferases (HATs) and 18 histone deacetylases (HDACs) have been identified to date, while the mechanistic details of substrate selection and site specificity of these enzymes remain unclear. The regulation of histone lysine acetylation status is impaired in the pathologies of cancer and other diseases and therefore, enzymes regulating histone lysine acetylation have become promising targets for anti-cancer drugs.Storage & StabilityThe antibody is delivered in lyophilized powder. Upon receipt, please centrifuge the antibody at 12,000 x g for 20 seconds, and then reconstitute it with the antibody stabilizer provided. The stabilizer contains PBS, 50% glycerol, and 0.01% sodium azide. Please store the antibody at -20°C. Avoid repeated freeze/thaw cycles. Antibody is stable for 12 months from date of reconstitution. Equilibrate the antibody to room temperature for 2 minutes and mix by gently pipetting before use.

References

• Xu, Qingqing, et al. “Hyper-acetylation contributes to the sensitivity of chemo-resistant prostate cancer cells to histone deacetylase inhibitor Trichostatin A.” Journal of Cellular and Molecular Medicine 22.3 (2018): 1909-1922.PMID: 29327812

**Research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.