Principle of Assay
This kit provides a simple method for distinguishing between healthy and apoptotic cells by detecting the changes in the mitochondrial transmembrane potential, based on a carbocyanine dye, JC-1. In healthy cells, this dye accumulates and aggregates in the mitochondria, where it forms bright red fluorescent agglomerates. Any event that dissipates the mitochondrial membrane potential prevents the accumulation of the JC-1 dye in the mitochondria and thus, the dye remains in the cytoplasm in its monomer form, leading to a shift from red (agglomerated JC-1, Ex/Em = 585/590 nm) to green fluorescence (JC-1 monomers, Ex/Em = 510/527 nm). JC-1 can be used as an indicator of mitochondrial potential in a variety of cell types, including myocytes and neurons, as well as in intact tissues and isolated mitochondria. This kit contains CCCP that causes an uncoupling of the proton gradient, which established during the normal activity of electron carriers in the electron transport chain, and thus, dissipates the mitochondrial electrochemical potential and may be used as a control that prevents JC-1 aggregation.
