| Background |
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 DAPI 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, and 4',6-Diamidino-2-Phenylindole dihydrochloride (4', 6-diamidino-2-phenylindole, DAPI) can enter the cells of late apoptotic and necrotic cells. It specifically binds to double-stranded DNA and generates intense blue fluorescence (Ex/Em: 358 nm/461 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. |
Reviews
There are no reviews yet.