Sonicated Chromatin prepared from untreated or 17beta-estradiol (E2) treated MCF7 cells was subjected to the ChIP procedure (see protocol) with ab412 to mono and dimethyl 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). 3 µl of ab412 and 4x10⁶ cells were used in each ChIP experiment.

RIP140 null adipocytes extracts were suspended in 250 µl of immunoprecipitation buffer (150 mM NaCl,50 mM Tris-HCl (pH 8.0), 1 mM EDTA, 0.2% (v/v) Nonidet P40, 2 mM PMSF, 0.1% (w/v) SDS and a protease-inhibitor cocktail. Protein extracts (200 µg) were incubated with antibodies to RIP140, 14-3-3, or PRMT1 overnight at 4°C, and precipitated with protein G–agarose beads for 1–2 hours. ab412 was then used (amongst other antibodies) in Western Blot. Reduced protein inputs were used for ectopic expression of RIP140 to avoid saturation.