Apo 3/7 HTS Assay Kit

Apoptosis is an evolutionarily conserved form of cell suicide, which follows a specialized cellular process. The central component of this process is a cascade of proteolytic enzymes called caspases. These enzymes participate in a series of reactions that are triggered in response to pro-apoptotic signals and result in cleavage of protein substrates, causing the disassembly of the cell. Caspases have been identified in organisms ranging from C. elegans to humans. The mammalian caspases play distinct roles in apoptosis and inflammation. In apoptosis, caspases are responsible for proteolytic cleavages that lead to cell disassembly (effector caspases), and are involved in upstream regulatory events (initiator caspases). An active caspase consists of two large (~20 kD) and two small (~10 kD) subunits to form two heterodimers which associate in a tetramer. As is common with other proteases, caspases are synthesized as precursors that undergo proteolytic maturation, either autocatalytically or in a cascade by enzymes with similar specificity. 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 C, actin a DNA-dependent protein kinase. This kit utilizes the quenched (z-DEVD)2-R110 peptide substrate for caspase 3/7 detection. The absorption and emission properties of the R110 dye are suppressed when attached to the z-DEVD peptide sequence. When R110 is cleaved away, by active caspase 3/7, from the quenching DEVD sequence, the free dye excites at 488 nm and emits at 515-530 nm. The Apo 3/7 HTS Assay Kit provides a homogenous platform that can be utilized for high throughput fluorescence plate reader applications. The reagent is directly added to the samples thus eliminating any wash steps.