Description

Description: Introduction Elabscience® Mitochondrial Membrane Potential Assay Kit (with JC-1) is developed to identify early apoptotic by detecting changes in mitochondrial membrane potential using JC-1 as a fluorescent probe. This kit provides carbonyl cyanide m-chlorophenylhydrazone (CCCP) [E-CK-A301C] to induce the decrease in mitochondrial membrane potential as a positive control reagent. Detection principle The decrease of mitochondrial membrane potential is a marker event in the early stage of apoptosis. It occurs before the appearance of nuclear apoptotic features (chromatin condensation and DNA fragmentation). Once the mitochondrial membrane potential collapses, apoptosis is irreversible. JC-1 is an ideal fluorescent probe widely used to detect mitochondrial membrane potential Δψm. In normal cells, the mitochondrial membrane potential is high, JC-1 aggregates in the mitochondrial matrix to form a polymer, which can produce red fluorescence. In the early stage of apoptosis, the mitochondrial membrane potential decreases, JC-1 can't accumulate in the mitochondrial matrix. When JC-1 is a monomer, it can produce green fluorescence. The decrease of cell membrane potential can be detected by the transition of JC-1 from red fluorescence to green fluorescence, and the transition of JC-1 fluorescence color can be used as an early detection indicator of cell apoptosis. The relative ratio of red and green fluorescence is commonly used to measure the ratio of mitochondrial depolarization. The maximum excitation wavelength of JC-1 monomer is 514 nm and the maximum emission wavelength is 529 nm; the maximum excitation wavelength of JC-1 polymer is 585 nm and the maximum emission wavelength is 590 nm. Components Materials not included Ultrapure water, vortex mixer, fluorescence microscope / laser scanning confocal microscope / flow cytometer Storage JC-1 Assay Buffer (10x) [E-CK-A301B] should be store at 2~8°C, other reagents should be stored at -20°C. JC-1 (500 x) [E-CK-A301A] and CCCP (10 mM) [E-CK-A301C] should be stored in dark. Avoid repeated freezing and thawing. Cautions 1. This kit is for research use only. 2. For your safety and health, please take safety precautions and follow the procedures of laboratory reagent operation. CCCP is an inhibitor of the mitochondrial electron transport chain, which is harmful to the human body. Please wear laboratory clothes and disposable gloves during operation, and avoid direct contact with the human body or inhalation of the body. 3. The validity period of this kit is 12 months. 4. JC-1 may coagulate or precipitate at lower temperatures. Please heat at 20~25°C water bath until it is completely dissolved. 5. IF the cells sensitive to pH changes, please use fetal bovine serum to replace JC-1 Assay Buffer for incubation or prolong observation time. 6. Fluorescent substances are prone to quenching. When performing fluorescent observations, shorten the observation time as much as possible and pay attention to store the sample with shading light.

Additional Text

2~8°C/-20°C, shading light. Refer to the Manual.