Caspase-8 / FLICE Fluorogenic Substrate
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- Application
- In vitro Assay (in vitro)
- Purpose
- Synthetic peptide substrate labeled at the carboxy end with AFC (7-amino-4-trifluoromethyl coumaride). Designed to measure caspase-8 (FLICE) activity in vitro.
- Sequence
- Ac-Ile-Glu-Thr-Asp-7-AFC, Ac-IETD-AFC
- Specificity
- Highly specific substrate for caspase-8. May be weak substrate for caspase-6.
- Purity
- > 98 % by HPLC
- Formula
- C31H38N5012F3
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- Application Notes
- For in vitro assays of caspase-8 activity. Can be used with purified or partially purified enzymes (~15 ng enzyme), or possibly with crude cell lysates (if the caspase-8 Inhibitor is included to determine background protease activity).
- Comment
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Fluorometer calibration: The fluorometer is calibrated using known concentrations of free AFC (Excitation = 400 nm, Emission = 505 nm) to generate a calibration curve of fluorescence versus µ-moles AFC. Samples: Can be either purified or partially purified enzyme preparations. Application to crude cell lysates has not been confirmed. If crude cell lysates are to be assayed, the non- specific protease background must be determined using our Caspase-8/FLICE Inhibitor, Z-IETD-FMK.
- Restrictions
- For Research Use only
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- Format
- Lyophilized
- Storage
- RT
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- Background
- Synthetic peptide substrate labeled at the carboxy end with AFC (7-amino-4-trifluoromethyl coumaride). Designed to measure caspase-8 (FLICE) activity in vitro. Caspase-8 (also known as FLICE, MACH or Mch5) is a member of the caspase family of cysteine proteases. Caspases play an important role in apoptosis signaling and effector mechanisms. Caspase-8 is most similar to caspase-10, both of which have death domain motifs. Caspase-8 appears to be physically associated with the signaling mechanism during Fas-mediated cell death and its association with Fas or tumor necrosis factor receptors via an interaction with FADD suggests it functions as an initiator rather than an effector of the cell death pathway. Thus, caspase-8 is an upstream activator in the protease cascade that proteolytically matures other caspases.
- Molecular Weight
- 729 Da
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