Caspase-9 Assay Kit, Fluorometric with Ac-LEHD-AFC substrate

Description

Caspase-9 Assay Kit, Fluorometric with Ac-LEHD-AFC substrate

General description

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 fluorescence microtiter plate reader.

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 an apoptotic sample with an uninduced control allows determination of the fold increase in Caspase-9 activity

Kit Components:

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

Additional information