Programmed cell death, or apoptosis, is the regulated elimination of cells that occurs naturally during cell development. Death domain proteins activate cysteine proteases of the ICE/CED-3 family. These proteases target various enzymes, initiating a proteolytic cascade of key proteins involved in cell function. This process, also known as the caspase cascade, ultimately leads to apoptosis (Figure 1). Other contributing or indicating factors in apoptosis are associated with a family of proteins related to Bcl2 and the collapse of the electrochemical gradient across the mitochondrial membrane.

Highly Specific Caspase Detection

Caspase 3—also known as CPP-32, Apopain, or Yama—is a key effector caspase in the apoptotic pathway. It is present in many different cell lineages and is responsible for the cleavage of a variety of molecules such as poly ADP-ribose polymerase (PARP), Protein Kinase Cd (PKC), actin, and DNA-dependent protein kinase. Our new Active Caspase Detection Kits utilize a polyclonal antibody against Caspase 3 and a secondary PElabeled goat anti-rabbit antibody to visualize the bound primary antibody. In comparison to other peptide-based (DEVD) assays which tend to crossreact with Caspase 7 and other caspases, our Active Caspase Detection Kits are highly specific for active human and murine Caspase 3. Both quantitative and qualitative results can be obtained with a flow cytometer, fluorescence microscope, or 96-well plate reader.

Convenient Mitochondrial Membrane Potential Detection

An early indicator of the initiation of cellular apoptosis is the collapse of the electrochemical gradient across the mitochondrial membrane. Our new Mitochondrial

Membrane Potential Detection Kit detects the loss in membrane potential by detecting the fluorescence of a unique dye, JC-1. In healthy cells, JC-1 enters the mitochondria and fluoresces red. In apoptotic cells, the mitochondrial membrane collapses and JC-1 disperses throughout the cell, while maintaining its green fluorescent monomeric form. This kit’s JC-1 reagent makes it easy to differentiate apoptotic cells from healthy cells. Apoptotic cells show primarily green fluorescence, while healthy cells show red and green fluorescence. Cells can be analyzed by flow cytometry, fluorescence plate reading, or fluorescence microscopy.

Extensive Line of Antibodies

For the detection of apoptosis-related proteins, we offer a new line of antibodies. Applications include Western blotting (Figure 2), immunoprecipitation, and immunohistochemistry.