Western blot analysis of Phospho-YAP1 (S127) on different lysates with Rabbit anti-Phospho-YAP1 (S127) antibody (ET1611-69) at 1/5;000 dilution. Lane 1: HeLa whole cell lysate Lane 2: HeLa treated with 100nM Calyculin A for 30 minutes whole cell lysate Lane 3: HeLa treated with 100nM Calyculin A for 30 minutes then treated with λpp for 1 hour whole cell lysate Lane 4: NIH/3T3 whole cell lysate Lane 5: NIH/3T3 treated with 100nM Calyculin A for 30 minutes whole cell lysate Lane 6: NIH/3T3 treated with 100nM Calyculin A for 30 minutes then treated with λpp for 1 hour whole cell lysate Lysates/proteins at 10 µg/Lane. Lane 7: C6 whole cell lysate Lane 8: C6 treated with 100nM Calyculin A for 30 minutes whole cell lysate Lane 9: C6 treated with 100nM Calyculin A for 30 minutes then treated with λpp for 1 hour whole cell lysate Predicted band size: 54 kDa Observed band size: 70 kDa Exposure time: 1 minute 6 seconds; 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 (ET1611-69) at 1/5;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.
Western blot analysis of SAV1 on different lysates with Rabbit anti-SAV1 antibody (HA722130) at 1/2;000 dilution and competitor's antibody at 1/2;000 dilution. Lane 1: HCT 116 cell lysate (20 µg/Lane) Lane 2: HeLa cell lysate (20 µg/Lane) Lane 3: 293T cell lysate (20 µg/Lane) Lane 4: HEK-293 cell lysate (20 µg/Lane) Lane 5: MDA-MB-231 cell lysate (20 µg/Lane) Lane 6: SW480 cell lysate (20 µg/Lane) Lane 7: BxPC-3 cell lysate (20 µg/Lane) Lane 8: 786-0 cell lysate (negative) (20 µg/Lane) Lane 9: A549 cell lysate (20 µg/Lane) Lane 10: NIH/3T3 cell lysate (20 µg/Lane) Lane 11: PC-12 cell lysate (20 µg/Lane) Lane 12: Mouse testis tissue lysate (40 µg/Lane) Lane 13: Rat testis tissue lysate (40 µg/Lane) Predicted band size: 45 kDa Observed band size: 45 kDa Exposure time: 3 minute4; ECL: Lane 1-13 (left): K1801; Lane 1-13 (right): K1802; 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 (HA722130) at 1/2;000 dilution and competitor's antibody at 1/2;000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50;000 dilution was used for 1 hour at room temperature.
