Caspase-1 Assay Kit, Fluorometric with Ac-YVAD-AFC substrate

Caspase-1 Assay Kit, Fluorometric with Ac-YVAD-AFC substrate

General description

Activation of ICE-family proteases/caspases initiates apoptosis or other cellular processes in mammalian cells. The Caspase-1/ICE Fluorometric Assay Kit provides a simple and convenient means for assaying the activity of caspases that recognize the sequence YVAD. The assay is based on detection of cleavage of substrate YVAD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin). YVAD-AFC emits blue light (400 nm); upon cleavage of the substrate by caspase-1 or related caspases, free AFC emits a yellow-green fluorescence (505 nm), which can be quantified using a fluorometer or a fluorescence microtiter plate reader. Comparison of the fluorescence of AFC from a treated sample with an untreated control allows determination of the fold increase in Caspase-1 activity.

Detection Method:         Fluorescence (400 nm excitation filter and 505 nm emission filter)

Species Reactivity:         Mammalian

Applications:                   Detect early/middle stages of apoptosis; differentiate apoptosis from necrosis

Storage temp.:               -20°C

Features & Benefits:

• Simple one-step procedure that takes ~1-2 hours
• Fast and convenient
• Comparison of the fluorescence of AFC from a treated sample with an untreated control allows determination of the fold increase in caspase-1 activity

Kit Components :

• Cell Lysis Buffer
• 2X Reaction Buffer
• YVAD-AFC (1mM)
• DTT (1 M)