Product Description
Size: 100 μl
Ab Type: Monoclonal
Reactivity: Eukaryotes
Immunogen: Acetylated H3 (Lys27)-KLH
Host: Mouse
Product DescriptionThis product is produced by immunizing mouses with a synthetic acetyl peptide corresponding to residues surrounding Lys27 of human histone H3. Antibodies are purified by protein G-conjugated agarose followed by acetylated histone H3 (Lys27) peptide affinity chromatography.
SpecificityThis antibody detects histone H3 only when it is acetylated at Lys27, but not the crotonylated, butyrylated, propionylated, and acetylated H3 peptide at Lys23, crotonylated or unmodified H3 peptide at Lys27, acetylated H3 peptide at Lys36.
Recommended ApplicationsChIP, IP. Not tested in other applications*IP=Immunoprecipitation ChIP=Chromatin precipitation
Recommended 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. Not yet tested in other applications.
A. Dot blotting analysis on indicated amount of acetylated H3 peptide at Lys27 (lane 1), crotonylated H3 peptide at Lys23 (lane 2), butyrylated H3 peptide at Lys23 (lane 3), propionylated H3 peptide at Lys23 (lane 4), acetylated H3 peptide at Lys23 (lane 5), crotonylated H3 peptide at Lys27 (lane 6), acetylated H3 peptide at Lys36 (lane 7) and unmodified H3 peptide at Lys27 (lane 8) using acetyl-histone H3 (Lys27) mouse mAb.
B. Sonicated chromatin prepared from 1x 105 HeLa cells treated with sodium butyrate was subjected to chromatin immunoprecipitation (ChIP) using either 4 μl Acetyl-Histone H3 (Lys27) mouse mAb or 1 μg of normal mouse 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.
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 Lys5, 9, 15 and 36; H2B is primarily acetylated at Lys5, 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.**Research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
