Western blotting analysis using FAS antibody (Cat#61981). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with FAS antibody (Cat#61981, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using NaQ™ ECL Substrate Kit (Cat#716).
Western blotting analysis using FAS antibody (Cat#61981). FAS expression in wild-type (WT) and FAS shRNA knockdown (KD) HT-1080 cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with FAS antibody (Cat#61981, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using NaQ™ ECL Substrate Kit (Cat#716).
Immunohistochemistry was performed on paraffin-embedded human tonsillitis using FAS antibody (Cat#61981, 1:200). Antigen retrieval was done in sodium citrate buffer (pH 6.0). DAB was used for detection, with hematoxylin counterstaining. Images were acquired using a Nikon Ci-L Plus microscope (40× objective). Scale bar: 25 μm.
Validation of FAS knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HT-1080 cells were stained with FAS antibody (Cat#61981, 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.
