Caspase-6 Substrate-pNA
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- Application
- In vitro Assay (in vitro), In vivo Studies (in vivo)
- Purpose
- Chromogenic paranitroanilide-peptide substrate for caspase-6 (Mch2).
- Sequence
- Acetyl-Val-Glu-Ile-Asp-pNA, Ac-VEID-pNA
- Specificity
- Substrate for caspase-3, 6.
- Purity
- > 98 % by HPLC
- Formula
- C28H40N6O11
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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-6 (Mch2). Release of free pNA is monitored by absorbance at 405 nm (epsilon=9,160 M^-1 cm^-1 ). Caspase-6 (also known as Mch2) is a member of the caspase family of cysteine proteases involved in apoptosis. It is a member of the Group III caspases (6, 8, and 9) which prefer the (L/V) EXD sequence as a substrate. Caspase-6 prefers a hydrophobic amino acid at P4, along with caspases-1 and -4, as opposed to the preference for Asp seen with caspases-2, -3, and -7. This is at odds with the gene sequence alignment that predicts Caspase-6 is more closely related to caspases-3 and -7 than to caspase-1. The preference by Caspase-6 for beta-branched amino acids in P4 fits well with the one known natural substrate, lamin A, and distinguishes it from caspases-1 and -4. Reconstitution experiments indicate that Caspase-6 activates caspases-3 and -7 and is therefore part of the proteolytic cascade that initiates apoptosis.
- Molecular Weight
- 636 Da
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