Product Description
The rabbit-derived antibody is purified with protein A-conjugated agarose followed by di-methylated histone H3 (Lys36) peptide affinity chromatography. It specifically recognizes histone H3 with di-methylation at Lys36.
Source/Purification
This product is produced by immunizing rabbits with a synthetic di-methyl peptide corresponding to residues surrounding Lys36 of human histone H3. Antibodies are purified by protein A-conjugated agarose followed by di-methylated histone H3 (Lys36) peptide affinity chromatography.
Specificity
H3
Recommended Applications
ELISA,Dot, WB, ICC, IHC; Not tested in other applications
*WB=Western blotting IHC=Immunohistochemistry ICC=Immunocytochemistry
Recommended antibody dilution
WB: 1:2000 IHC: 1:1000 ICC: 1:200
Important
For Western blotting, incubate membrane with diluted antibody in 5% nonfat milk, 1 x TBS, 0.1% Tween-20 fortwo hours at room temperaturewith gentle shaking.
Use at an assay dependent concentration. Optimal dilutions/concentrations should be determined by the end user.Not yet tested in other applications.
![Anti-di-methyl-Histone H3 (Lys36) rabbit pAb]( "Anti-di-methyl-Histone H3 (Lys36) rabbit pAb")
| A. Dot blotting analysis on indicated amount of di-methylated H3 peptide at Lys36 (lane 1), mono-methylated H3 peptide at Lys36 (lane 2), tri-methylated H3 peptide at Lys36 (lane 3) and unmodified H3 peptide at Lys36 (lane 4) using di-methyl-histone H3 (Lys36) rabbit pAb. | B. Western blotting analysis on 30 μg of crude proteins from HeLa, 3T3 and H1299 whole cell lysates using di-methyl-histone H3 (Lys36) rabbit pAb (1:2000). |
![Anti-di-methyl-Histone H3 (Lys36) rabbit pAb]( "Anti-di-methyl-Histone H3 (Lys36) rabbit pAb")
| C. Immunohistochemical analysis of paraffin-embedded mouse colon tissue, labeling H3K36me2 with PTM-624 at 1/1000 dilution. Nuclear staining on epithelial cells of mouse colon was observed. Counter stained with hematoxylin. |
![Anti-di-methyl-Histone H3 (Lys36) rabbit pAb]( "Anti-di-methyl-Histone H3 (Lys36) rabbit pAb")
| D. Immunocytochemistry analysis of Hela (Human cervix adenocarcinoma cell line), labeling H3K36me2 with PTM-624 at 1/200 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Nuclei were stained with DAPI (blue). |
Scientific Description
The ε-amino lysine acetylation 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) is impaired in the pathologies of cancer and other diseases and therefore, enzymes regulating histone lysine methylation have become promising targets for anti-cancer drugs.
Storage & Stability
Store product at -20oC. Avoid repeated freeze/thaw. Antibody is supplied in PBS with 50% glycerol and 0.01% sodium azide.Stable for 12 months from date of receipt.
**Research purposes only.Not intended to be used for therapeutic or diagnostic purposes in humans or animals.