Western blotting analysis using JUN antibody (Cat#63465). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with JUN antibody (Cat#63465, 1:2,500) and HRP-conjugated goat anti-mouse secondary antibody (Cat#101, 1:20,000) respectively. Image was developed using NaQ™ ECL Substrate Kit(Cat#716).
Western blotting analysis using JUN antibody (Cat#63465). JUN expression in wild type (WT) and JUN shRNA knockdown (KD) C2C12 cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with JUN antibody (Cat#63465, 1:5,000) and HRP-conjugated goat anti-mouse secondary antibody (Cat#101, 1:20,000) respectively. Image was developed using NaQ™ ECL Substrate Kit (Cat#716).
Immunocytochemical staining of HT-1080 cells with JUN antibody (Cat#63465, 1:1,000). Nuclei were stained blue with DAPI; JUN 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 JUN expression in HT-1080 cells using JUN antibody (Cat#63465, 1:2,000). Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 0.5% Triton X-100 in 1x PBS. Green, isotype control; red, JUN.
Validation of JUN knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) C2C12 cells were stained with JUN antibody (Cat#63465, 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.
