Mitochondrial membrane potential assay kit with JC-1

The Mitochondrial Apoptosis Detection Kit provides a simple, fluorescent-based method for distinguishing between healthy and apoptotic cells by detecting the loss of the mitochondrial transmembrane potential (ΔΨ). Disruption of the mitochondrial transmembrane potential is one of the earliest intracellular events that occur in the induction of apoptosis. The kit utilizes the unique lipophilic cationic dye (5, 5’, 6, 6’-tetrachloro-1, 1’, 3, 3’- tetraethyl- benzimidazolylcarbocyanine iodide, commonly known as JC-1) for detection. In healthy cells, the negative charge established by the intact mitochondrial membrane potential allows the lipophilic dye, bearing a delocalized positive charge, to enter the mitochondrial matrix where it accumulates. With the increasing of the concentration of JC-1, it aggregates and become fluorescent red. When the mitochondrial potential collapses in apoptotic cells, JC-1 just exists as monomers and do not accumulate within the mitochondria. When dispersed in this manner, JC-1 remains in the cytoplasm in a green fluorescent monomeric form. The fluorescent signals can be easily detected by fluorescence microscopy using a band-pass filter (detects FITC and rhodamine), or analyzed by flow cytometry using FITC channel for green monomers (Ex/Em =488/530+30 nm) and PI channel for red aggregates (Ex/Em =488/590+42 nm).