Rb2 p130 r27020

Cells were lysed in a buffer containing 0.1% Triton (Bio-Rad, Irvine, CA, USA) for 30 min in ice. Then, 20 μg of each lysate was electrophoresed in a polyacrylamide gel and electroblotted onto a nitrocellulose membrane. We used the following primary antibodies: RB1 (AV33212) and GAPDH (G8795) were from Sigma-Aldrich (MO, USA), RB2/P130 (R27020) was from BD Biosciences (San Jose, CA, USA), p27^KIP1 (3686) was from Cell Signaling, p107 (sc-318), p53 (sc-126), and p21^CIP1 (sc-397) were from Santa Cruz Biotechnology (Dallas, TX, USA), and p16^INK4A (ab54210) was from Abcam (Cambridge, UK). Immunoreactive signals were detected with a horseradish peroxidase-conjugated secondary antibody (ImmunoReagents, Raleigh, NC, USA) and reacted with ECL plus reagent (Merck Millipore). All of the antibodies were used according to the manufacturer’s instructions. The mean value was quantified densitometrically using Quantity One 1-D analysis software (Bio-Rad).

Cells were lysed in a buffer containing 0.1% Triton (Bio-Rad, Hercules, CA, USA) for 30 min in ice; 20 μg of each lysate was electrophoresed in a polyacrylamide gel and electroblotted onto a nitrocellulose membrane. We used the following primary antibodies: RB1 (AV33212) and GAPDH (G8795) from Sigma-Aldrich (St. Louis, MO, USA); RB2/P130 (R27020) from BD Biosciences (Allschwil, Switzerland); and P27^KIP1 (3686) from Cell Signaling Technology (Danvers, MA, USA), while TP53 (sc-126) and P21^CIP1 (sc-397) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and P^16INK4A (ab54210) was obtained from Abcam (Cambridge, UK). Immunoreactive signals were detected with a horseradish peroxidase conjugated secondary antibody (ImmunoReagents, Raleigh, NC, USA) and reacted with ECL plus reagent (Merck Millipore, Burlington MA, USA). All antibodies were used according to manufacturer’s instructions. The mean value was quantified densitometrically using Quantity One^® 1-D analysis software (Bio-Rad, Hercules, CA, USA).

Cells were lysed in a buffer containing 0.1% Triton (Bio-Rad, CA, USA) for 30 min in ice. 20 μg of each lysate was electrophoresed in a polyacrylamide gel and electroblotted onto a nitrocellulose membrane. We used the following primary antibodies: RB1 (AV33212) and GAPDH (G8795) were from Sigma-Aldrich (MO, USA), RB2/P130 (R27020) was from BD Biosciences (CA, USA), p27^KIP1 (3686) was from Cell Signaling (MA, USA), while p107 (sc-318), p53 (sc-126), and p21^CIP1 (sc-397) were obtained from Santa Cruz Biotechnology (CA, USA), and p16^INK4A (ab54210) was from ABCAM (Cambridge, UK). Immunoreactive signals were detected with a horseradish peroxidase-conjugated secondary antibody (ImmunoReagents, NC, USA) and reacted with ECL plus reagent (Merck Millipore, MA, USA). All of the antibodies were used according to the manufacturer’s instructions. The mean value was quantified densitometrically using Quantity One 1-D analysis software (Bio-Rad, CA, USA).