Anti ctip

Cell extracts were obtained by lysing cells for 15 min at 4C in buffer containing 50 mM Tris-HCl (pH 8.0), 300 mM NaCl, 0.4% NP-40, 10 mM MgCl2 and 5 mM DTT, supplemented with protease and phosphatase inhibitors (Halt Protease & Phosphatase Inhibitor Cocktail; ThermoFisher Scientific). After centrifugation (10,000 x g, 20 min), supernatants were diluted (v/v) in 50 mM Tris-HCl (pH 8.0), 0.4% NP-40 and 5 mM DTT. Proteins were separated by SDS-PAGE and immunoblotted with the following antibodies at dilutions recommended by the manufacturer: anti-pan-actin (MAB1501; Millipore), anti-ARTEMIS (#13381; Cell Signaling Technology), antimCherry (GTX128509; GeneTex), anti-CtIP (A300–488A; Bethyl), anti-FEN1 (A300–255A; Bethyl), anti-KAP1 (A300–274A; Bethyl), anti-DNA ligase3 (A301–636A; Bethyl), anti-MRE11 (A303–998A; Bethyl), anti-MUS81 (ab14387; Abcam), anti-PNKP (A300–257A; Bethyl), anti-SETX (A301–104A; Bethyl), anti-TDP1 (H00055775-A01; Abnova Corporation), anti-atubulin (T5168; Sigma-Aldrich), anti-XPF (A301–315A; Bethyl), anti-XPG (A301–484A; Bethyl), and anti-XRCC1 (A300–065A; Bethyl). Immunoblotting was revealed by chemiluminescence using a ChemiDoc MP Imaging System (Bio-Rad).