Phycoerythrin pe conjugated goat anti mouse igg

HAP1 cells kindly provided by Thjin R. Brummelkamp (Netherlands Cancer Institute, Amsterdam, The Netherlands) were cultured in Iscove’s modified Dulbecco’s medium containing 10% fetal calf serum with necessary selection antibiotics [18 (link)]. Mouse monoclonal anti-CD59 (clone 5H8) and anti-DAF (clone IA10) were used as primary antibodies [22 (link)]. Phycoerythrin (PE)-conjugated goat anti-mouse IgG (Biolegend) was used as a secondary antibody. The bacterial pore-forming toxin proaerolysin and its variant fluorescent-labeled aerolysin (FLAER) were obtained from Protox Biotech [23 (link)].

Human embryonic kidney 293 (HEK293) cells were cultured in Dulbecco’s Modified Eagle medium (DMEM) supplemented with 10% fetal calf serum. The appropriate antibiotic concentrations were used where necessary: puromycin (1 μg/ml), hygromycin (400 μg/ml), and streptomycin (100 U/ml)/penicillin (100 μg/ml). Mouse monoclonal anti-CD59 (clone 5H8), which was kindly provided from Taroh Kinoshita (Osaka University), and anti-M2 Flag antibodies (Sigma, St. Louis, MO, USA) were used as primary antibodies, whereas phycoerythrin (PE)-conjugated goat anti-mouse IgG (Biolegend, San Diego, CA, USA) was used as the secondary antibody. 1-(2-Deoxy-2-fluoro-β-D-arabinofuranosyl)-5-iodo-2,4 (1H,3H)-pyrimidinedione (FIAU) and 4-NPP were purchased from Sigma.