Western blotting analysis using Cdk4 antibody (Cat#67244). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with Cdk4 antibody (Cat#67244,1:5,000) and HRP-conjugated goat anti rabbit secondary antibody (Cat#201,1:50,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Western blotting analysis using Cdk4 antibody (Cat#67244). Cdk4 expression in wild type (WT) and Cdk4 shRNA knockdown (KD) HT-1080 cells with 30 μg of total cell lysates. GAPDH serves as a loading control. The blot was incubated with Cdk4 antibody (Cat#67244, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:50,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Immunocytochemical staining of HepG2 cells with CDK4 antibody(Cat#67244, 1:1,000). Nuclei were stained blue with DAPI; CDK4 was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: Medium. Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 0.5% Triton X-100 in 1x PBS. Scale bar, 20 μm.
Immunocytochemical staining of HeLa cells using CDK4 antibody (Cat#67244, 1:1,000), Top panel: wild-type (WT); Bottom panal: CDK4 shRNA knockdown (KD). Nuclei were stained blue with DAPI; CDK4 was stained magenta with Alexa Fluor® 647. Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 0.5% Triton X-100 in 1x PBS. Scale bar, 20 μm.
Flow cytometric analysis of Cdk4 expression in HepG2 cells using Cdk4 antibody (Cat#67244,1:2,000). Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 100% Methanol, chill on -20℃ before use. Green, isotype control; red, Cdk4.
Validation of Cdk4 knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with Cdk4 antibody (Cat#67244, 1:2,000) and analyzed using BD flow cytometer. Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 100% Methanol, chill on -20℃ before use.
