Caspase-9 Fluorometric Assay Kit

Activation of ICE-family proteases/caspases initiates apoptosis in mammalian cells. The Caspase-9 Fluorometric Assay Kit provides a simple and convenient means for assaying the activity of caspases that recognize the sequence LEHD. The assay is based on detection of cleavage of substrate LEHD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin). LEHD-AFC emits blue light (λmax = 400 nm); upon cleavage of the substrate by caspase-9 or related caspases, free AFC emits a yellow-green fluorescence (λmax = 505 nm), which can be quantified using a fluorometer or fluorecence microtiter plate reader.

• Detection method: fluorecence (400 nm excitation filter and 505 nm emission filter) • Species reactivity: Mammalian • Kit size: Convenient sizes (25, 100, 200, 400 assays) • Applications: Detect early/middle stages of apoptosis; differentiate apoptosis from necrosis.

• Simple one-step procedure; takes 1-2 hours • Fast and convenient • Comparison of the fluorescence of AFC from an apoptotic sample with an uninduced control allows determination of the fold increase in caspase-9 activity.

Apoptosis Assay Kit

none

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

-20°C

gel pack

6-12 months

see datasheet

Detect early/middle stages of apoptosis; differentiate apoptosis from necrosis.

Cell and tissue lysates

Biovision supplies other types of Assays as 1.

Human and some mouse caspases are active in apoptosis and cell death and even in necrosis and inflammation. CASP Gene and orthologous enzymes have been identifies successfully in the signal transduction cascade and pathways.