ApoStat Intracellular Caspase Detection (FITC-VAD-FMK)

Detection of Caspase Activation in Jurkat Cells. Human Jurkat leukemic T cells were cultured in the absence (green) or presence (purple) of 1 µM staurosporine for 3.5 hours and then stained with ApoStat (Catalog # FMK012) for 30 minutes. The cells were harvested, washed, and assayed by flow cytometry to determine the percentage of cells expressing active caspases.

ApoStat is designed to identify and quantify caspase activity in apoptotic cells by flow cytometry. Cells undergoing apotosis are irreversibly labeled with a cell permeable, FITC-conjugated pan-caspase inhibitor (ApoStat). Any unbound reagent diffuses out of the cell and is washed away. Cells are then analyzed by flow cytometry for the presence of bound reagent. Increased fluorescence is an indicator of caspases activity within cells. The detection of active caspases by flow cytometry is a rapid method for identifying cells undergoing apoptosis.

ApoStat Apoptosis Detection Kit

Label:FITC
Testing Format: Flow Cytometry
Sample Type: Cultured cells
Size: 100 Tests

Caspases are a family of cytosolic aspartate-specific cysteine proteases involved in the initiation and execution of apoptosis. They are expressed as latent zymogens and are activated by an autoproteolytic mechanism or by processing by other proteases (frequently other caspases). Human caspases can be subdivided into three functional groups: cytokine activation (caspase-1, -4, -5, and -13), apoptosis initiation (caspase-2, -8, -9, -and -10), and apoptosis execution (caspase-3, -6, and -7).

Caspases are regulated by a variety of stimili, including APAF1, CFLAR/FLIP, NOL3/ARC, and members of the inhibitor of apoptosis (IAP) family such as BIRC1/NAIP, BIRC2/cIAP-1, BIRC3/cIAP-2, BIRC4/XIAP, BIRC5/Survivin, and BIRC7/Livin. IAP activity is modulated by DIABLO/SMAC or PRSS25/HTRA2/Omi. Cell-permeable and irreversible peptide inhibitors are also available for different caspases.