This gene encodes a member of the cysteine-aspartic acid protease (caspase) family. Sequential activation of caspases plays a central role in the execution-phase of cell apoptosis. Caspases exist as inactive proenzymes which undergo proteolytic processing at conserved aspartic residues to produce two subunits, large and small, that dimerize to form the active enzyme. This protein can undergo autoproteolytic processing and activation by the apoptosome, a protein complex of cytochrome c and the apoptotic peptidase activating factor 1; this step is thought to be one of the earliest in the caspase activation cascade. This protein is thought to play a central role in apoptosis and to be a tumor suppressor. Alternative splicing results in multiple transcript variants.

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Immunocytochemical staining of HeLa cells using Cleaved Caspase-9 antibody , Top panel: wild-type (WT); Bottom panal: Cleaved Caspase-9 shRNA knockdown (KD). Nuclei were stained blue with DAPI; Cleaved Caspase-9 was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm. Permeabilization: Triton.

Flow cytometric analysis of cleaved Caspase-9 expression in HepG2 cells using cleaved Caspase-9 antibody . Green, isotype control; red, cleaved Caspase-9.

Immunocytochemical staining of HepG2 cells with cleaved Caspase-9 antibody . Nuclei were stained blue with DAPI; cleaved Caspase-9 was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: Medium. Scale bar, 20 μm.

Western blotting analysis using cleaved Caspase-9 antibody . Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with cleaved Caspase-9 antibody and HRP-conjugated goat anti-rabbit secondary antibody respectively. Image was developed using FeQ™ ECL Substrate Kit .

Western blotting analysis using cleaved Caspase-9 antibody . Cleaved Caspase-9 expression in wild type (WT) and cleaved Caspase-9 shRNA knockdown (KD) HeLa cells with 30 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with cleaved Caspase-9 antibody and HRP-conjugated goat anti-rabbit secondary antibody respectively. Image was developed using FeQ™ ECL Substrate Kit .

Validation of cleaved Caspase-9 knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with cleaved Caspase-9 antibody and analyzed using BD flow cytometer.

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