Western blot analysis of Phospho-RPA32/RPA2 (T21) on different lysates with Rabbit anti-Phospho-RPA32/RPA2 (T21) antibody (ET1611-18) at 1/1,000 dilution. Lane 1: HeLa whole cell lysate Lane 2: HeLa treated with 1uM Camptothecin for 1 hour whole cell lysate Lane 3: HEK-293 whole cell lysate Lane 4: HEK-293 treated with UV for 20 minutes, then recover 2 hours whole cell lysate Lane 5: HEK-293 treated with UV for 40 minutes, then recover 2 hours whole cell lysate Lane 6: HeLa whole cell lysate Lane 7: HeLa treated with UV for 40 minutes, then recover 2 hours whole cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 29 kDa Observed band size: 29/35 kDa Exposure time: 1 minute; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1611-18) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.