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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.
|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|
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.
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