Flow cytometric analysis of Cyclin D1 expression in C2C12 cells using Cyclin D1 antibody (Cat#69541, 1:2,000). Green, isotype control; red, Cyclin D1.
Immunocytochemical staining of C2C12 cells with Cyclin D1 antibody (Cat#69541, 1:1,000). Nuclei were stained blue with DAPI; Cyclin D1 was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: High. Scale bar: 20 μm.
Western blotting analysis using anti-Cyclin D1 antibody (Cat#69541). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with anti-Cyclin D1 antibody (Cat#69541, 1:10,000) 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 anti-Cyclin D1 antibody (Cat#69541). Cyclin D1 expression in wild type (WT) and Cyclin D1 shRNA knockdown (KD) HeLa cells with 30 μg of total cell lysates. β-Tubulin serves as a loading control. The blot was incubated with anti-Cyclin D1 antibody (Cat#69541, 1:10,000) and HRP-conjugated goat anti rabbit secondary antibody (Cat#201, 1:50,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Validation of Cyclin D1 knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with anti-Cyclin D1 antibody (Cat#69541, 1:2,000) and analyzed using BD flow cytometer.
Immunocytochemical staining of HeLa cells using anti-Cyclin D1 antibody (Cat#69541, 1:1,000), Top panel: wild-type (WT); Bottom panal: Cyclin D1 shRNA knockdown (KD). Nuclei were stained blue with DAPI; Cyclin D1 was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm. Permeabilization: Triton.