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Synthetic peptide corresponding to Human Histone H3 aa 1-100 (asymmetric di methyl R17) conjugated to Keyhole Limpet Haemocyanin (KLH).
(Peptide available as
The nuclear hormone receptor co-activator CARM1 has the potential to methylate histone H3 at arginine residues in vitro. The methyltransferase activity of CARM1 is necessary for its co-activator functions in transient transfection assays. However, the role of this methyltransferase in vivo is unclear, given that methylation of arginines is not easily detectable on purified histones. This antibody recognizes methylated arginine 17 (R17) of histone H3, the major site of methylation by CARM1. Bauer et al (2001) have shown by using this antibody that methylated R17 exists in vivo. Chromatin immunoprecipitation analysis shows that R17 methylation on histone H3 is dramatically upregulated when the estrogen receptor-regulated pS2 gene is stimulated by estradiol and TPA. Coincident with the appearance of methylated R17, the CARM1 methyltransferase is found associated with the histones on the pS2 gene. Together these results demonstrate that the CARM1 methyltransferase is recruited to an active promoter and that CARM1-mediated methylation of histone H3 at R17 takes place in vivo during this active state.
Our Abpromise guarantee covers the use of ab8284 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ChIP/Chip||Use at an assay dependent dilution.|
|PepArr||Use a concentration of 0.002 - 0.0002 µg/ml.|
|IHC-P||Use at an assay dependent dilution.|
|ICC/IF||Use a concentration of 0.1 µg/ml.|
|ChIP||Use at an assay dependent dilution.|
|Dot Blot||Use at an assay dependent dilution.|
|IP||Use at an assay dependent dilution.|
|WB||1/1000 - 1/2000. Can be blocked with Histone H3 (asymmetric di methyl R17) peptide (ab16935).|
All batches of ab8284 are tested in Peptide Array against peptides to different Histone H3 modifications. Six dilutions of each peptide are printed on to the Peptide Array in triplicate and results are averaged before being plotted on to a graph. Results show strong binding to Histone H3 - asymmetric di methyl R17 peptide (ab16935), indicating that this antibody specifically recognises the Histone H3 - asymmetric di methyl R17 modification.
Total U2OS cell extract was western blotted using the anti-Me-R17H3 antibody. The asterisk indicates methylated histone H3. The left panel shows presence of core histones (indicated on the left) by Coomassie Blue staining. Molecular weights are indicated on the right.
ab8284 was used in immunohistochemistry with paraffin embedded sections of human tonsil, using DAB as a chromogen (brown). Counterstaining of nuclei was performed with haemotoxylin (blue).
Staining is seen confined to the nucleus.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"