Caspase-2,3 Substrate -pNA
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- Application
- In vitro Assay (in vitro), In vivo Studies (in vivo)
- Purpose
- Chromogenic paranitroanilide-peptide substrate for caspase-2 (ICH-1).
- Sequence
- Acetyl-Val-Asp-Val-Ala-Asp-pNA, Ac-VDVAD-pNA
- Specificity
- Substrate for caspase-2, 3.
- Purity
- > 97 % by HPLC
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- Application Notes
- Assay of caspase activity in cell extracts.
- Comment
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Soluble in DMSO and aqueous buffers. We recommend preparing a stock solution in DMSO, and diluting into aqueous buffer shortly prior to use.
1. Lyse cells in 50 mM Tris-HCl, pH 7.5, 0.3% NP-40, 1.0 mM DTT, at a density of 2 X 10^6 /mL.
2. Assay 0.01 mL cell lysate in a final volume of 0.1 mL. Assay buffer is cell lysis buffer containing 0.2 mM substrate.
3. Incubate at 37°C for 0-3 hr. Take periodic readings of absorbance at 405 nm. - Restrictions
- For Research Use only
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- Format
- Lyophilized
- Storage
- RT
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- Background
- Chromogenic paranitroanilide-peptide substrate for caspase-2 (ICH-1). Release of free pNA is monitored by absorbance at 405 nm (epsilon=9,160 M^-1 cm^-1 ). 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-pNA substrate is excellent for studying Caspase-2.
- Molecular Weight
- 679 Da
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