Abcam's Caspase 3 (active) FITC Staining Kit provides a convenient means for sensitive detection of activated caspase 3 in living cells. The assay utilizes the caspase 3 inhibitor, DEVD-FMK, conjugated to FITC (FITC-DEVD-FMK) as a marker. FITC-DEVD-FMK is cell permeable, non-toxic, and irreversibly binds to activated caspase 3 in apoptotic cells. The FITC label allows detection of activated caspase-3 in apoptotic cells directly by fluorescence microscopy, flow cytometry, or fluorescence plate reader. Visit our FAQs page for tips and troubleshooting.
Activation of caspases plays a central role in apoptosis.
The caspase family of cysteine proteases play a key role in apoptosis. Caspase 3 (also known as CPP32, YAMA and apopain) is the most extensively studied apoptotic protein among caspase family members. Caspase 3 is synthesized as an inactive pro enzyme that is processed in cells undergoing apoptosis by self proteolysis and/or cleavage by other upstream proteases (e.g. Caspases 8, 9 and 10).
The processed form of Caspase 3 consists of large (17kD) and small (12kD) subunits which associate to form an active enzyme. Caspase 3 is cleaved at Asp28 - Ser29 and Asp175 - Ser176. The active Caspase 3 proteolytically cleaves and activates other caspases (e.g. Caspases 6, 7 and 9), as well as relevant targets in the cells (e.g. PARP and DFF). Alternative splicing of this gene results in two transcript variants which encode the same protein.
In immunohistochemical studies Caspase 3 expression has been shown to be widespread but not present in all cell types (e.g. commonly reported in epithelial cells of skin, renal proximal tubules and collecting ducts). Differences in the level of Caspase 3 have been reported in cells of short lived nature (eg germinal centre B cells) and those that are long lived (e.g. mantle zone B cells).
Caspase 3 is the predominant caspase involved in the cleavage of amyloid beta 4A precursor protein, which is associated with neuronal death in Alzheimer's disease.
Caspase 3 in Jurkat cells following 24 hour exposure to 2 uM Camptothecin (CPT; ab120115) with or without 20 nM caspase inhibitor Z-VAD(OMe)-FMK (zVAD; ab120487). Background signal subtracted, duplicates; +/- SD.