Caspase-6 Substrate-pNA

Produktnummer ABIN924949

Kurzübersicht für Caspase-6 Substrate-pNA (ABIN924949)

Applikation: In vitro Assay (in vitro), In vivo Studies (in vivo)

Produktdetails

Verwendungszweck: Chromogenic paranitroanilide-peptide substrate for caspase-6 (Mch2).

Sequenz: Acetyl-Val-Glu-Ile-Asp-pNA, Ac-VEID-pNA

Spezifität: Substrate for caspase-3, 6.

Reinheit: > 98 % by HPLC

Formel: C28H40N6O11

Anwendungsinformationen

Applikationshinweise: Assay of caspase activity in cell extracts.

Kommentare:

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.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Format: Lyophilized

Lagerung: RT

Antigendetails

Hintergrund: 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.

Molekulargewicht: 636 Da