Western blotting analysis using NELFB antibody (Cat#61643). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with NELFB antibody (Cat#61643, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Western blotting analysis using NELFB antibody (Cat#61643). NELFB expression in wild-type (WT) and NELFB shRNA knockdown (KD) HSHC cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with NELFB antibody (Cat#61643, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Immunocytochemical staining of HepG2 cells with NELFB antibody (Cat#61643, 1:1,000). Nuclei were stained blue with DAPI; NELFB 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 HAP1 cells using NELFB antibody (Cat#61643, 1:1,000), Top panel: wild-type (WT); Bottom panal: NELFB shRNA knockdown (KD). Nuclei were stained blue with DAPI; NELFB 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.
Validation of NELFB knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HSHC cells were stained with NELFB antibody (Cat#61643, 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.
