Western blot analysis of Phospho-Rb (S795) on different lysates with Rabbit anti-Phospho-Rb (S795) antibody (HA722764) at 1/2;000 dilution. Lane 1: HeLa cell lysate (20 µg/Lane) Lane 2: HeLa treated with 100ng/mL Nocodazole for 18 hours cell lysate (20 µg/Lane) Lane 3: HeLa treated with 100ng/mL Nocodazole for 18 hours cell lysate; then the membrane treated with λpp for 1 hour (20 µg/Lane) Predicted band size: 106 kDa Observed band size: 106 kDa Exposure time: 3 minutes; ECL: K1801; 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 (HA722764) at 1/2;000 dilution was 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.
Western blot analysis of Phospho-Rb (S807) on different lysates with Rabbit anti-Phospho-Rb (S807) antibody (ET1602-36) at 1/2;000 dilution. Lane 1: K-562 cell lysate (no heat) (20 µg/Lane) Lane 2: K-562 cell lysate (20 µg/Lane) Lane 3: K-562 cell lysate (no heat); treated with λpp for 1 hour (20 µg/Lane) Lane 4: K-562 cell lysate; treated with λpp for 1 hour (20 µg/Lane) Predicted band size: 106 kDa Observed band size: 106 kDa Exposure time: 2 minutes; 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 (ET1602-36) at 1/2;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 Phospho-Rb (S780) on different lysates with Rabbit anti-Phospho-Rb (S780) antibody (HA721513) at 1/1;000 dilution. Lane 1: Jurkat serum starved for 44 hours whole cell lysate Lane 2: Jurkat serum starved for 24 hours then 10% FBS incubated for 20 hours whole cell lysate Lane 3: Jurkat serum starved for 44 hours then treated with λpp for 1 hour whole cell lysate Lane 4: Jurkat serum starved for 24 hours then 10% FBS incubated for 20 hours then treated with λpp for 1 hour whole cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 106 kDa Observed band size: 106 kDa Exposure time: 1 minute 59 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 (HA721513) 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.
Western blot analysis of Phospho-Rb (S795) on different lysates with Rabbit anti-Phospho-Rb (S795) antibody (HA722764) at 1/2;000 dilution and competitor's antibody at 1/2;000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 100ng/mL Nocodazole for 18 hours cell lysate Lane 3: U-2 OS cell lysate Lane 4: U-2 OS treated with 50ng/mL Nocodazole for 8 hours cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 106 kDa Observed band size: 106 kDa Exposure time: Lane 1-4 (left): 40 seconds; Lane 1-4 (right): 1 minute; ECL: 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 (HA722764) 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.
