Scrape each plate of cells into 1 ml of ice-cold EBC lysis buffer (50 mM Tris (pH 8), 120 mM NaCl, 0.5% NP-40, 50 μg/ml leupeptin, 10 μg/ml aprotinin, 50 μg/ml phenylmethylsulfonyl fluoride ...

Pellet cells at 300g at 4 °C for 10 min in a centrifuge. 3| Resuspend the cell pellet in 5 ml of ice-cold NP-40 lysis buffer. Incubate on ice for 5 min. Pellet the nuclei at 120g at 4 °C for 10 ...