Western blotting analysis using calpastatin antibody (Cat#63547). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with calpastatin antibody (Cat#63547, 1:5,000) and HRP-conjugated goat anti-mouse secondary antibody (Cat#101, 1:20,000) respectively. Image was developed using FeQTM ECL Substrate Kit (Cat#226).
Western blotting analysis using calpastatin antibody (Cat#63547). Calpastatin expression in wild type (WT) and calpastatin (CAST) shRNA knockdown (KD) HeLa cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with calpastatin antibody (Cat#63547, 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 HeLa cells using calpastatin antibody (Cat#63547, 1:1,000), Top panel: wild-type (WT); Bottom panal: CAST shRNA knockdown (KD). Nuclei were stained blue with DAPI; Calpastatin 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 Calpastatin knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with Calpastatin antibody (Cat#63547, 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.
