Caspase-1 / ICE Fluorogenic Substrate 2

Catalog No. ABIN924939

Quick Overview for Caspase-1 / ICE Fluorogenic Substrate 2 (ABIN924939)

Application: In vitro Assay (in vitro)

Product Details

Purpose: Peptide substrate labeled at the carboxy end with AFC (7-amino-4-trifluoromethyl coumarin). Designed to measure Caspase-1 or Caspase-4 activity in vitro.

Sequence: Ac-Tyr-Val-Ala-Asp-AFC, Ac-YVAD-AFC

Specificity: Serves as a substrate for Caspase-1 and as a weak substrate for Caspase-4. Not cleaved by Caspases-2, -3, -6, or -7.

Application Details

Application Notes: For in vitro assays of Caspase-1 and Caspase-4 activities. Can be used with purified or partially purified enzymes, or possibly with crude cell lysates (if the Caspase-1/ICE Inhibitor 2 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 caspase 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 specific Caspase-1/ICE Inhibitor 2.

Restrictions: For Research Use only

Handling

Storage: RT

Target Details

Background: Peptide substrate labeled at the carboxy end with AFC (7-amino-4-trifluoromethyl coumarin). Designed to measure Caspase-1 or Caspase-4 activity in vitro. Interleukin-1beta Converting Enzyme (ICE), now termed Caspase-1, is a cytoplasmic cysteine protease that cleaves inactive 31 kDa pro-IL-1beta to generate the active 17.5 kDa proinflammatory cytokine IL-1beta, the predominant form of IL-1 produced by human monocytes. This cytokine has been implicated in the pathogenesis of several diseases such as rheumatoid arthritis, inflammatory bowel disease, and septic shock. Caspase-1/ICE mRNA is found in a variety of cells such as peripheral blood monocytes, peripheral blood lymphocytes, peripheral blood neutrophils, and resting and activated peripheral blood T lymphocytes. The tissue distribution of Caspase-1/ICE suggests that the enzyme may have other substrates in addition to IL-1beta. Current hypotheses suggest that Caspase-1/ICE is able to cause apoptosis as well as activate inflammation in animal cells. Experiments have shown that Caspase-1/ICE has sequence homology with other mammalian apoptosis genes and that activation of Caspase-1/ICE or other ICE-related proteases (caspases)is required for anti-Fas mAb-induced apoptosis.

Molecular Weight: 720 Da