All lanes : Anti-TRRAP antibody - ChIP Grade (ab73546) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Nuclear LysateLane 2 : Jurkat (Human T cell lymphoblast-like cell line) Nuclear Lysate (ab14844)Lysates/proteins at 10 µg per lane.SecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
Chromatin was prepared from HeLa cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 5µg of ab73546 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach for both the active and heterochromatic loci). Primers are located in the first kb of the transcribed region.