Key features and details
- Assay type: Quantitative
- Platform: Microplate reader
- Assay time: 1 hr 30 min
- Sample type: Adherent cells, Suspension cells
Product nameCaspase-10 Inhibitor Assay Kit
See all Caspase-10 kits
Sample typeAdherent cells, Suspension cells
Assay time1h 30m
Abcam's Caspase 10 Inhibitor Drug Detection Kit provides an effective means for screening caspase inhibitors using fluorometric methods. The assay utilizes synthetic peptide substrate AEVD-AFC (AFC, 7-amino-4-trifluoromethyl coumarin). Active caspase 10 cleaves the synthetic substrate to release free AFC which can then be quantified by fluorometry. Compounds to be screened can directly be added to the reaction and the level of inhibition of caspase 10 activity can be determined by comparison of the fluorescence intensity in samples with and without the testing inhibitors.
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Caspases have been shown to play a crucial role in apoptosis induced by various deleterious and physiologic stimuli. Inhibition of caspases can delay apoptosis, implicating a potential role in drug screening efforts.
Storage instructionsStore at -20°C. Please refer to protocols.
Components 100 tests Active Caspase-10 1 x 100 units Caspase Inhibitor, Z-VAD-FMK 1 x 10µl Caspase Substrate AEVD-AFC 1 x 0.5ml DTT 1 x 100µl Reaction Buffer 1 x 10ml
RelevanceCaspases are a family of intracellular proteases that mediate cell death and are the principal effectors of apoptosis. Caspase 10 (Mch4, ICE-LAP4, FLICE2) plays an important role in apoptosis induced by a variety of inducers such as TNF alpha. It is a large prodomain caspase classified together with caspases 2, 8, and 9 as a signaling caspase. Four isoforms of caspase 10 (caspase 10a, 10b, 10c, and 10d) having the same prodomain but different mature large and small subdomain, have been described. Caspase 10 contains two death domains (DED) involved in linking to the death effector domain of the adapter protein FADD and recruiting the complex to TNFR1 and Fas. The inactive procaspase 10 is variably expressed in many tissues and cell lines as a cytosolic protein. The mature form of caspase 10 comprises two subunits, p23/p17 (splice isoforms) and p12. Caspase 10 can cleave and activate caspases 3, 4, 6, 7, 8, and 9. This is followed by cleavage of numerous key proteins, including the nuclear protein PARP.
- Apoptotic protease MCH 4
- CASP 10
Caspase activity (RFU) in presence of 0µM - 40µM of z-VAD-FMK (generic caspase inhibitor), assessed using AEVD-AFC as caspase 10 substrate and following Caspase 10 Inhibitor Drug Detection Kit (ab102498) protocol. Lanes:
1.- Background Control: no Caspase 10, no z-VMAD-FMK
2.- Positive Inhibition Control: no Caspase 10, +z-VAD-FMK
3.- Positive Control: + Caspase 10, no z-VAD-FMK
4.- Caspase 10 + 2.5µM z-VAD-FMK
5.- Caspase 10 + 10µM z-VAD-FMK
6.- Caspase 10 + 40µM z-VAD-FMK
ab102498 has not yet been referenced specifically in any publications.