Caspase 3/7 and PI Apoptosis Double Staining Kit

Caspase 3 (CPP32/ Apopain) plays a significant role in the initiation of apoptosis, it has been widely accepted as an indicator of apoptosis. Caspase 3 has a specific substrate polypeptide recognition sequence, namely aspartic acid-glutamate-valine-aspartic acid (DEVD) . The Caspase 3/7 and PI Apoptosis Double Staining kit is used for apoptosis detection by measuring the activity of Caspase 3/7 and the integrity of the cell membrane. Caspase 3/7 Green uses the Caspase 3/7 recognition sequence (DEVD) coupled to a high-affinity DNA fluorescent dye, which has cell membrane permeability and can penetrate the cell membrane into the cytoplasm. The Caspase 3/7 Green reagent itself is non-fluorescent and has a charge repulsion effect with DNA. When cells undergo apoptosis, Caspase 3/7 Green, as a substrate, is cleaved by activated Caspase 3/7 and releases a high-affinity DNA dye. After binding to DNA, it generates strong fluorescence (Ex/Em: 488 nm/525 nm, flow cytometry recommended FITC channel) . When cells enter the late stage of apoptosis or necrosis, the integrity of the cell membrane is lost. Propidium Iodide (PI) can enter the cells of late apoptotic and necrotic cells, specifically bind to double-stranded DNA and produce intense red fluorescence (Ex/Em:535 nm/617 nm) . When used in combination with Caspase 3/7 Green, it can simultaneously distinguish the activity of Caspase 3/7 enzymes in necrotic cells and apoptotic cells.