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This antibody will be of central importance in analysing the methylation status of chromatin and transcription factors. Since it recognises monomethyl arginine only it can be used in parallel with ab413 (detects dimethyl arginine only) and ab412 (detects both monomethyl and dimethyl arginine) to monitor the exact modification status.
Our Abpromise guarantee covers the use of ab415 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent dilution.|
|ChIP||Use 5µl for 106 cells.|
Sonicated Chromatin prepared from untreated or 17beta-estradiol (E2) treated MCF7 cells was subjected to the ChIP procedure (see protocol) with ab415 to mono methyl Arginine.
Immunoprecipitated chromatin was analysed in the proximal region of the estrogen-responsive pS2 promoter (as shown above) and quantified by real-time PCR (values are % of inputs). The primers are designed to follow the nucleosome E (including the Estrogen Responsive Element ERE). 5
We looked at chicken chromatin and then analysed 4 active genes (AG1-4), 2 inactive genes (IG1-2) as well as heterochromatin (Het). A "Yes" indicates that the antibody immunoprecipitated material well above background and should be regarded as a positive result.
Chromatin source: Chicken erythrocytes. Chromatin was formaldehyde crosslinked and then sonicated. The chromatin was then purified on a CsCl gradient.
ChIP: The Abcam protocol from the website was employed. Approximately 4-5 ug of each antibody was used with 30-40 ug of chromatin per ChIP.
PCR: An Applied Biosystems ABI 7000 Real-time PCR instrument was used. Taqman probe technology was used.
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