Product Description
Size: 100 μl
Ab Type: Polyclonal
Reactivity: Eukaryotes
Immunogen: Mono-methylated H3 (Lys56)-KLH
Host: Rabbit
Product DescriptionThis product is produced by immunizing rabbits with a synthetic mono-methyl peptide corresponding to residues surrounding Lys56 of human histone H3. Antibodies are purified by protein A-conjugated agarose followed by mono-methylated histone H3 (Lys56) peptide affinity chromatography.
SpecificityThis antibody detects histone H3 only when it is mono-methylated at Lys56, but not the acetylated, di-, tri-methylated or unmodified H3 peptide at Lys56.
Recommended ApplicationsChIP, IP. Not tested in other applications*IP=Immunoprecipitation ChIP=Chromatin precipitation
Recommended antibody dilutionIP: 1:25 ChIP: 1:50 (2 μl of antibody for 1x 105 HeLa)
Important NotesUse at an assay dependent concentration. Optimal dilutions/concentrations should be determined by the end user.
A. Dot blotting analysis on indicated amount of mono-methylated H3 peptide at Lys56 (lane 1), acetylated H3 peptide at Lys56 (lane 2), di-methylated H3 peptide at Lys56 (lane 3), tri-methylatedH3 peptide at Lys56 (lane 4) and unmodified H3 peptide at Lys56 (lane 5) using mono-methyl-histone H3 (Lys56) rabbit pAb.
B. Sonicated chromatin prepared from 1x 105 HeLa cells was subjected to chromatin immunoprecipitation (ChIP) using either serial dilutions of Mono-methyl-Histone H3 (Lys56) 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 Promoter, RPL30 Exon 3, MyoD1 Exon 1 and β-globin. Data are presented as enrichment of each IPed sample relative to total amount of input chromatin at each amplicon.
Scientific DescriptionThe ε-amino lysine methylation of proteins is an important reversible modification controlling protein activity. The amino-terminal tails of core histones undergo lysine methylation in multiple sites, termed as “histone code” or “epigenetic code”. Lysine methylation in core histones is a major determinant for the formation of active and inactive regions of the genome and therefore plays vital roles in multiple cellular events. In most species, lysine methylation occurs primarily on histones H3 (Lys4, 9, 27, 36, 79), and H4 (Lys20) and has been implicated in both transcriptional activation and silencing. Methylation in histones modulated by specific histone methylases (HMTs) and histone demethylases (HDMs). The level of methyllysineis found impaired in the pathologies of cancers and other diseases, therefore, enzymes regulating histone lysine methylation 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.
