Western blotting analysis using STAT3 antibody (Cat#63413). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with STAT3 antibody (Cat#63413, 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). STAT3, signal transducer and activator of transcription 3.
Western blotting analysis using STAT3 antibody (Cat#63413). STAT3 expression in wild type (WT) and STAT3 shRNA knockdown (KD) HeLa cells with 20 μg of total cell lysates. β-Tubulin serves as a loading control. The blot was incubated with STAT3 antibody (Cat#63413, 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 STAT3 antibody (Cat#63413, 1:1,000). Nuclei were stained blue with DAPI; STAT3 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 STAT3 expression in HepG2 cells using STAT3 antibody (Cat#63413, 1:2,000). Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 0.5% Triton X-100 in 1x PBS. Green, isotype control; red, STAT3.
Validation of STAT3 knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with STAT3 antibody (Cat#63413, 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.
