Caspase 3 Activity Assay

Benefits
Specifically detect caspase 3 activity: Detect natural and recombinant human caspase 3 activity - not other caspases - using the caspase 3-specific anti-CPP32 monoclonal capture antibody
Achieve highly sensitive detection of caspase 3 activity: Detect extremely low levels of caspase 3 activity in lysates using the kit’s fluorescent substrate in an immunosorbent enzyme assay
Screen multiple samples simultaneously: Take advantage of the kit’s convenient 96-well microplate format when screening multiple samples
Store lysates for extended periods: Since the same buffer serves as both lysis and reaction buffers, you can immediately assay lysates or store them for subsequent analysis
Background Information
The initiation of all intracellular events seems to be characterized by the activation of specific proteases called caspases (cysteinyl-aspartic-acid-proteases). Biological function of Caspase 3 (CPP32, Yama, Apopain) activation seems to play a key role during the apoptotic process. Activated CPP32 is responsible for the cleavage, and therefore the breakdown, of several cellular components related to DNA repair and regulation. Cleavage sites of different caspases, including CPP32, are described.
Contents
Coating Buffer, 10x, 5 ml
Anti-Caspase-3, 20x, 500 µl monoclonal antibody from mouse
Blocking Buffer, ready-to-use, 25 ml
Incubation Buffer, 5x, 50 ml
DTT, 100x, 400 µl (1 M) stock solution
Substrate Solution Ac- DEVD-AFC, 20x, 650 µl (1 mM) stock solution in DMSO
AFC, 150 µl (1 mM) Stock Solution in DMSO
Positive Control, 250 µl lysate from apoptotic U937 cells
Microplate, 12 modules of 8 wells each
Adhesive Plate Cover Foils, 2 sheets
Application
Samples suitable for the Caspase 3 Activity Assay are:
Lysates of apoptosis-induced cells
Recombinant caspase 3 (CPP32)
The Caspase 3 Activity Assay can be used to:
Detect and quantify in vitro caspase 3 activity in cell lysates
Analyze caspase 1, 2, 3, or 7 activity by simply omitting the capture-antibody step
Perform inhibitor studies for Caspase 3
This kit is intended as a tool for life-science research to further the understanding of apoptosis.