Immunofluorescence - Anti-Phospho-Rb (Thr373) Rabbit mAb [54C33H63]

Sample: HeLa cells

The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.5% Triton X-100 for 10 minutes and then blocked with 5% BSA in 0.1% PBS-Tween for 0.5 hours.

Primary antibodies: R015398 at 1:100 dilution and α-tubulin Mouse Monoclonal Antibody (Cat. No. LF209) at 1:100 dilution

Secondary antibodies: Goat anti-Rabbit (488) at 1:1,000 dilution (shown in green) and Goat anti-Mouse (555) at 1:1,000 dilution (shown in red)

Nuclei were stained with DAPI (shown in blue).

Western Blot - Anti-Phospho-Rb (Thr373) Rabbit mAb [54C33H63]

All lanes: R015398 at 1:10,000 dilution

Lane 1: HCT116 (Human colorectal carcinoma epithelial cell) whole cell lysates

Lysates/proteins at 10 µg per lane.

Secondary antibody: Goat Anti-Rabbit IgG (H+L), HRP Conjugated (Cat. No. LF102) at 1:5,000 dilution

Predicted band size: 106 kDa

Observed band size: 110 kDa

Developed using the ECL technique (Cat. No. SQ201).