Key features and details
- Assay type: Enzyme activity
- Platform: Microplate reader, Fluor. microscope, Flow cyt.
- Assay time: 2 hr
- Sample type: Adherent cells, Suspension cells
Product nameCleaved Caspase-3 Staining Kit (FITC)
See all Caspase-3 kits
Sample typeAdherent cells, Suspension cells
Assay typeEnzyme activity
Assay time2h 00m
Cleaved Caspase-3 Staining Kit (FITC) ab65613 provides a convenient means for sensitive detection of activated Caspase-3 in living cells.
The cleaved caspase 3 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 / cleaved caspase-3 in apoptotic cells directly by fluorescence microscopy, flow cytometry, or fluorescence plate reader.
PlatformMicroplate reader, Fluor. microscope, Flow cyt.
Storage instructionsStore at -20°C. Please refer to protocols.
Components 100 tests FITC-DEVD-FMK 1 x 100µl Wash Buffer 2 x 100ml Z-VAD-FMK 1 x 10µl
RelevanceThe 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.
- Apopain precursor
Caspase-3 activation was measured by flow cytometry, using ab65613, in cells that were left alone or pretreated with QVD.oph and then treated with I-CRP (1.25 U/ml) or a drug that induces DNA strand breaks (100 µM) for 24 h, data was then analyzed and graphed.
ab65613 has been referenced in 13 publications.
- Zheng JJ et al. Novel role of PAF1 in attenuating radiosensitivity in cervical cancer by inhibiting IER5 transcription. Radiat Oncol 15:131 (2020). PubMed: 32471508
- Martínez-Torres AC et al. Chitosan gold nanoparticles induce cell death in HeLa and MCF-7 cells through reactive oxygen species production. Int J Nanomedicine 13:3235-3250 (2018). PubMed: 29910612
- Martínez-Torres AC et al. IMMUNEPOTENT CRP induces cell cycle arrest and caspase-independent regulated cell death in HeLa cells through reactive oxygen species production. BMC Cancer 18:13 (2018). Flow Cyt ; Human . PubMed: 29298674
- Våtsveen TK et al. Artesunate shows potent anti-tumor activity in B-cell lymphoma. J Hematol Oncol 11:23 (2018). PubMed: 29458389
- Lu L et al. Gold-chrysophanol nanoparticles suppress human prostate cancer progression through inactivating AKT expression and inducing apoptosis and ROS generation in vitro and in vivo. Int J Oncol 51:1089-1103 (2017). PubMed: 28849003