Neurological Diseases
Cancer Research
Sperm Viability
Drugs Development
Understanding the mechanisms of programmed cell death (also known as apoptosis) is an important aspect in the research of e.g., tissue homeostasis, tumour diseases, or the toxicity and efficacy of drugs, among others.
Why is Annexin V Staining used to detect apoptosis?
The human vascular anticoagulant, Annexin V is a 35-36KDa Ca2+ dependent phospholipids binding protein that has a high affinity for PS. In healthy cells, PS is located on the cytoplasmic surface of the plasma membrane.
However, during early apoptosis, phosphatidylserine (PS), normally located on the cytoplasmic surface of the cell membrane, becomes exposed to the extracellular environment, causing changes in the membrane asymmetry and permeability.
Mitochondrial Assays: detection of early apoptotic cells
The dysfunction of the mitochondria is a special feature of earlier stages of apoptosis. MitoStep apoptosis kits measure changes in the mitochondrial membrane potential with a solution of the cationic cyanine dye DiIC1(5).
In healthy cells, especially in mitochondria with active membrane potentials, the dye accumulates. During apoptosis, the DiIC1(5) fluorescence intensity decreases and thus there is an increase of cells with lower DiIC1(5) fluorescence.
MitoStep Kits:
Depolarization of the submitochondria and thus changes in the mitochondrial membrane during the early stages of apoptosis have been described. This is the reason why mitochondrial uptake of cationic dyes is a possible source of fluorescence variation.
DiIC1(5)-stained cells can be visualized by excitation with a 633 nm laser and emission at 658 nm, and combined with FITC or PE staining.
