Western blotting analysis using Annexin V antibody (Cat#69585). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with Annexin V antibody (Cat#69585,1:10,000) and HRP-conjugated goat anti rabbit secondary antibody (Cat#201,1:50,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Western blotting analysis using Annexin V antibody (Cat#69585). Annexin V expression in wild type (WT) and Annexin V shRNA knockdown (KD) HeLa cells with 30 μg of total cell lysates. β-Tubulin serves as a loading control. The blot was incubated with Annexin V antibody (Cat#69585,1:10,000) and HRP-conjugated goat anti rabbit secondary antibody (Cat#201,1:50,000) respectively. Image was developed using FeQ™ ECL Substrate Kit (Cat#226).
Immunocytochemical staining of HT-1080 cells with Annexin V antibody (Cat#69585, 1:1,000). Nuclei were stained blue with DAPI; Annexin V was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: High. 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 Annexin V expression in HT-1080 cells using Annexin V antibody (Cat#69585,1:2,000). Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 100% Methanol, chill on -20℃ before use. Green, isotype control; red, Annexin V.
Validation of Annexin V knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with Annexin V antibody (Cat#69585, 1:2,000) and analyzed using CytoFLEX. Fixative solution: 4% Paraformaldehyde in 1x PBS; Permeabilization solution: 100% Methanol, chill on -20℃ before use.
