Western blotting analysis using phospho-HSF1 (Ser303/Ser307) antibody (Cat#63389). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with phospho-HSF1 (Ser303/Ser307) antibody (Cat#63389, 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 phospho-HSF1 (Ser303/Ser307) antibody (Cat#63389). Phospho-HSF1 (Ser303/Ser307) expression in wild type (WT) and HSF1 shRNA knockdown (KD) 293T cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with phospho-HSF1 (Ser303/Ser307) antibody (Cat#63389, 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). HSF1, heat shock transcription factor 1.
Immunocytochemical staining of HepG2 cells with phospho-HSF1 (Ser303/Ser307) antibody (Cat#63389, 1:1,000). Nuclei were stained blue with DAPI; Phospho-HSF1 (Ser303/Ser307) 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 Phospho-HSF1 (Ser303/Ser307) expression in HepG2 cells using Phospho-HSF1 (Ser303/Ser307) antibody (Cat#63389, 1:2,000). Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 0.5% Triton X-100 in 1x PBS. Green, isotype control; red,Phospho-HSF1 (Ser303/Ser307).
