Anti histone h3 dimethyl k27 antibody

Chromatin was extracted from naive CD4⁺Foxp3⁻ T cells (1 × 10⁶ cells) polarized for 24–72 h under various conditions after fixation with formaldehyde. Anti-Histone H3ac (39139, 7 μl), anti-Histone H4ac (39243, 6 μl), anti-Histone H3 (trimethyl K9) (61013, 7 μl), anti-Histone H3 (dimethyl K27) antibody (39245, 5 μl; all from Active Motif), anti-HDAC1 (ab7028, 5 μg), anti-Smad3 (ab28379, 5 μg), anti-Histone H3 (trimethyl K27) (ab6002, 5 μl), anti-Histone H3 (acetyl K27) (ab4729, 3 μl), anti-Histone H3 (acetyl K9) (ab4441, 4 μl; all from Abcam), anti-dimethyl-Histone H3 (diMe-Lys9) (D5567, 5 μl; Sigma- Aldrich), anti-p50 (sc-1192X, 5 μg), anti-p300 (sc-585X, 5 μg), anti-STAT6 (SC-981X, 5 μg), control mouse IgG (sc-2025, 5 μg; all from Santa Cruz Technology) and purified goat IgG (02–6202, 5 μg; Life Technologies) were used for the immunoprecipitation of chromatin with an EZ ChIP kit (17–371; EMD Millipore) according to the manufacturer's instructions. The precipitated DNA was measured by real-time PCR using SsoAdvanced Universal SYBR Green Supermix (172–5275; Bio-Red). Real-time PCR primer sequences are listed in Supplementary Table 2. Data are presented as relative binding based on normalization to input DNA50 (link).