Caspase-2, 3 Fluorogenic Substrate
-
- Application
- In vitro Assay (in vitro)
- Purpose
- Peptide substrate labeled at the carboxy end with AFC (7-amino-4-trifluoromethyl coumarin). Designed to measure Caspase-2 activity in vitro.
- Sequence
- Ac-Val-Asp-Val-Ala-Asp-AFC, Ac-VDVAD-AFC
- Specificity
- Highly specific substrate for Caspase-2. Will also serve as a substrate for Caspases-3 and -7.
-
-
- Application Notes
- For in vitro assays of Caspase-2 activity, or Caspases-3 and -7 although the Caspase-3 Fluorogenic Substrate (Ac-DEVD-AFC, Cat. No. AC-003) is preferable for these two enzymes. The Caspase-2,3 Fluorogenic Substrate can be used with purified or partially purified enzymes or possibly with crude cell lysates (if the Caspase-2,3 Inhibitor is included to determine background protease activity).
- Comment
-
Fluorometer calibration: The fluorometer is calibrated using known concentrations of free AFC (Excitation = 400 nm, Emission = 505 nm) to generate a standard 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-2,3 Inhibitor.
- Restrictions
- For Research Use only
-
- Storage
- RT
-
- Background
- Peptide substrate labeled at the carboxy end with AFC (7-amino-4-trifluoromethyl coumarin). Designed to measure Caspase-2 activity in vitro. Caspase-2 (also known as ICH-1) is a member of the caspase family of cysteine proteases involved in apoptosis. It is a member of the Group II caspases, along with caspases-3 and - 7. Group II caspases prefer peptides of the DEXD-type as substrates. However, unlike caspases-3 and -7, Caspase-2 requires a P5 amino acid in the peptide for efficient cleavage. The similar substrate specificities of the Group II caspases suggests that their roles in cells are at least overlapping, if not completely redundant. The requirement for a fifth amino acid in substrates for Caspase-2 means that the DEVD-type, while serving as substrates for caspases-3 and -7, do not work with Caspase-2. For this reason, the Ac-VDVAD-AFC substrate is excellent for studying Caspase-2.
- Molecular Weight
- 770 Da
-
You are here:
