Western blotting analysis using P4HB antibody (Cat#62838). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with P4HB antibody (Cat#62838, 1:5,000) and HRP-conjugated goat anti-mouse secondary antibody (Cat#101, 1:20,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Western blotting analysis using P4HB antibody (Cat#62838). P4HB expression in wild type (WT) and P4HB shRNA knockdown (KD) HT-1080 cells with 30 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with P4HB antibody (Cat#62838, 1:5,000) and HRP-conjugated goat anti-mouse secondary antibody (Cat#101, 1:20,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Immunocytochemical staining of HepG2 cells with P4HB antibody (Cat#62838, 1:1,000). Nuclei were stained blue with DAPI; P4HB 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.
Flow cytometric analysis of P4HB expression in HepG2 cells using P4HB antibody (Cat#62838, 1:2,000). Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 0.5% Triton X-100 in 1x PBS. Green, isotype control; red, P4HB.
Validation of P4HB knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HT1080 cells were stained with P4HB antibody (Cat#62838, 1:2,000) and analyzed using BD flow cytometer. Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 0.5% Triton X-100 in 1x PBS.
