| Product Name | Mitochondrial Membrane Potential Assay Kit (JC-1) |
| Catalogue No | KTA4001 |
| Applications notes | Mitochondrial Membrane Potential Assay Kit (JC-1) 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. |
| Kit components | • JC-1 Stain(100×) • CCCP (50 mM) • PBS (10×) |
| Features & Benefits | • Measure mitochondrial membrane potential as an indicator of cell health. • Use JC-1, a fluorescent, lipophilic, cationic dye. • Red fluorescence indicates healthy mitochondria and green fluorescence indicates mitochondria in poor health. • Optimized for flow cytometery or fluorescent microscopy. • Contain CCC (Potent mitochondrial oxidative phosphorylation uncoupler). |
| Usage notes | • JC-1 Stain is difficult to prepare due to its low solubility. Make sure JC-1 Stain is completely thawed and warmed to room temperature before using. • JC-1 Stain is light senstive. Incubations need to be done in the dark. |
| Storage | Refer to list of materials supplied for storage conditions of individual components. Stable for at least 12 months at recommended temperature from date of shipment. |
| Shipping | Gel pack with blue ice. |
| Background | Mitochondria, the site of most energy production in eukaryotic cells, have a double membrane structure: an outer membrane and a folded inner membrane. Across the inner membrane of intact mitochondria there is a voltage gradient with the inside negative and the outside positive. Disruption of the mitochondrial transmembrane potential is one of the earliest intracellular events that occur following induction of apoptosis. Same time it can reduce O2 to produce O2-, which is the main part of the respiration electron transport chain to produce O2-. The activity of Complex I can not only reflect the status of responsibility electron transfer chain (ETC), but also reflect the production of reactive oxygen species (ROS) .monitored to check the efficacy of therapeutics such as insulin and sulfonylureas in type 2 diabetics. |
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