The MMP Orange Reagent Kit assesses the state of cellular mitochondrial health by detecting changes in the mitochondrial membrane potential (MMP). The MMP Orange Reagent diffuses across the intermembrane space and accumulates in proportion to the mitochondrial membrane potential and is used as a measurement of the electrochemical gradient across the mitochondrial membrane (m). In a healthy cell, there is a strong proton motive force driving ATP synthase and metabolism. A drop in MMP signals a disruption in mitochondrial integrity and possibly early apoptosis. Changes in MMP are used as metabolic markers associated with a variety of disease states. The assay utilizes two control compounds: Trifluoromethoxy carbonylcyanide phenylhydrazone (FCCP), a compound known for depolarizing the mitochondrial membrane (causing a decrease in MMP reagent fluorescence), and Oligomycin A, a compound known for polarizing the mitochondrial membrane (causing an increase in MMP reagent fluorescence). These reagents have been validated for use with the IncuCyte® S3 Live-Cell Analysis System for Metabolism and enables the real-time evaluation of mitochondrial membrane potential. The MMP Orange Reagent Kit is not compatible with IncuCyte instruments configured with a Red/Green Optical Module, e.g. Cat. No. 4647. The IncuCyte MMP Orange Reagent Kit is intended for use with the IncuCyte® Cell-by-Cell Analysis Software Module (Cat. No. 9600-0031).
Determine the state of mitochondria in live cells. Addition of MMP Orange Reagent to HeLa cells enable kinetic monitoring of the mitochondrial membrane potential state (A). Histograms reveal shifts in mean fluorescent intensity for control and drug treated wells at 0 and 40hrs, highlighting the change in mitochondrial polarization. Leverage label-free cell counting with the Cell-by-Cell Analysis Software for real -time data of cell proliferation (C) derived from HD phase/fluorescent images (D).