Abbkine 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.
Abbkine 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.
Product Properties
Kit components
• JC-1 Stain
• CCCP (10 mM)
• Assay Buffer (5×)
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 instructions
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.
Precautions
The product listed herein is for research use only and is not intended for use in human or clinical diagnosis. Suggested applications of our products are not recommendations to use our products in violation of any patent or as a license. We cannot be responsible for patent infringements or other violations that may occur with the use of this product.
Additional Information
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.
Anthocyanins have been reported to have effective chemopreventive activity. Lycium ruthenicum Murray is rich in anthocyanins and exhibits many biological activities. The purpose of this study was to investigate the effects and possible biological mechanism of the main anthocyanin monomer (Pt3G) of Lycium ruthenicum Murray on prostate cancer DU-145 cells. The cell proliferation was detected by methyl thiazolyl tetrazolium assay. The cell apoptosis rates were assessed by flow cytometric analysis and TUNEL assay. The expressions of apoptosis related proteins were evaluated by western blotting. Our data demonstrated that Pt3G inhibited cell proliferation, induced apoptosis and promoted cell cycle arrest at the S phase in a concentration-dependent manner (0, 100, 200 and 400 μg mL−1). Furthermore, it was shown that Pt3G decreased the mitochondrial membrane permeability through regulating the expressions of Bax and Bcl-2. Western blot analysis indicated that Pt3G significantly increased the expression of PTEN and then activated the PI3K/Akt-mediated caspase 3 pathway. In addition, our results also suggested that Pt3G activated the PTEN gene to induce the apoptosis of DU-145 cells by stimulating the overproduction of ROS. To sum up, these results indicate that Pt3G inhibits cell proliferation and induces apoptosis through the ROS/PTEN/PI3K/Akt/caspase 3 signaling pathway in prostate cancer DU-145 cells. Therefore, Pt3G of Lycium ruthenicum Murray may be a potential anti-proliferative agent for the prevention or treatment of prostate cancer.
To compare embryonic developmental competence and clinical outcomes of oocytes matured in vivo (IVF oocytes) and those matured in vitro (IVM oocytes) from the same IVM/IVF cycles, and to analyze the clinical efficiency of a melatonin-supplemented in vitro maturation system combined with a modified IVM/IVF protocol.
Key Findings
There were no significant differences in fertilization or blastocyst formation rates between the IVF and IVM groups, whereas the cleavage rate was significantly higher in the IVF versus IVM group (100% vs 93.4 ± 10.9%, p = 0.03). There were no significant differences in the clinical pregnancy, implantation or live birth/ongoing pregnancy rates between the two groups. The cumulative clinical pregnancy and ongoing pregnancy/live birth rate per pick-up oocyte in the IVM/IVF treatment cycles were 68.2% (15/22) and 54.5% (12/22), respectively. The reactive oxygen species and Ca2+ levels were significantly increased, and mitochondrial membrane potential was significantly decreased, in IVM compared with IVF oocytes.