Sigma-Aldrich Mitochondrial Membrane Potential Kit, sufficient for 500 fluorometric tests (microplate readers)

The Sigma-Aldrich Mitochondrial Membrane Potential Kit is an advanced tool designed to measure the mitochondrial membrane potential (MMP) in mammalian cells. This kit uses JC-10, a superior alternative to JC-1, to assess the loss of mitochondrial membrane potential during apoptosis. The kit is suitable for fluorometric assays using microplate readers, enabling precise monitoring of MMP collapse, which plays a crucial role in the apoptotic process.

Key Features and Benefits

• Accurate Measurement of MMP:
  The Mitochondrial Membrane Potential Kit is ideal for measuring the collapse of mitochondrial membrane potential (MMP), a key indicator of apoptosis. The kit allows the detection of mitochondrial dysfunction by using JC-10, a lipophilic cationic dye that enters mitochondria and exhibits a color shift in response to changes in membrane potential.
• Superior JC-10 Dye:
  Unlike JC-1, which can suffer from poor water solubility at higher concentrations, JC-10 has improved solubility and greater sensitivity in aqueous solutions. This makes it a better option for studying MMP without the precipitation issues that sometimes arise with JC-1.
• Fluorescent Detection:
  JC-10 forms red-fluorescent aggregates (λex = 540/λem = 590 nm) in cells with a polarized mitochondrial membrane, allowing detection of healthy cells with intact MMP. In apoptotic cells, MMP collapse results in a shift of JC-10 to its green fluorescent monomeric form (λex = 490/λem = 525 nm), making it easy to monitor the apoptotic process.
• Versatile and Efficient:
  This kit is designed for screening apoptosis inhibitors and activators with the help of microplate readers, allowing high-throughput screening and quantitative analysis of MMP in a variety of cell types.
• Cost-Effective and Reliable:
  The kit offers 500 fluorometric tests, providing a cost-effective solution for labs that need to perform multiple assays over time. The reliable performance of the kit makes it a trusted choice for studying apoptosis and mitochondrial function.

Applications

The Sigma-Aldrich Mitochondrial Membrane Potential Kit is suitable for various applications, including:

• Monitoring Apoptosis:
  The kit is essential for assessing mitochondrial dysfunction during apoptosis and can be used to evaluate the loss of MMP, which precedes the release of cytochrome c and the activation of the apoptotic cascade.
• Screening Apoptosis Inhibitors and Activators:
  This kit can be used in drug discovery and pharmacological studies to identify inhibitors or activators of apoptosis by assessing changes in MMP.
• Mitochondrial Function Studies:
  The kit is ideal for research related to mitochondrial dysfunction, oxidative stress, and cellular health. It can be used to evaluate the effects of various treatments on mitochondrial membrane integrity and cell viability.
• High-Throughput Screening:
  Suitable for high-throughput assays in a 96-well plate format, making it an efficient tool for large-scale screening and analysis of apoptotic pathways in mammalian cells.

Principle

This kit uses JC-10 dye, which is similar to JC-1, but with improved aqueous solubility. Here's how the assay works:

1. Healthy Cells (Polarized MMP):
   In cells with intact mitochondrial membrane potential, JC-10 forms red-fluorescent aggregates within the mitochondria. This fluorescence is detectable at 590 nm after excitation at 540 nm.
2. Apoptotic Cells (MMP Collapse):
   In apoptotic cells, the collapse of MMP prevents JC-10 from aggregating in the mitochondria, resulting in a shift to the green fluorescent monomeric form. This fluorescence is detectable at 525 nm after excitation at 490 nm.
3. Fluorescent Detection:
   The shift in fluorescence (from red to green) is a key indicator of the loss of MMP, which can be quantified using a microplate reader with fluorescent detection capabilities.

Kit Components

The Mitochondrial Membrane Potential Kit includes the following components:

• JC-10 Dye
• Dilution Buffers
• Positive Control (Apoptotic Cells)
• Microplate Reader-Compatible Fluorescent Detection Reagents
• User Manual

Specifications

Storage and Handling

• Storage:
  The kit should be stored at -20°C upon receipt. Ensure the components are protected from light to maintain their effectiveness.
• Handling:
  Always thaw the reagents at room temperature and avoid repeated freeze-thaw cycles to maintain the integrity of the components.

The Sigma-Aldrich Mitochondrial Membrane Potential Kit provides a highly sensitive, reliable, and easy-to-use method for measuring mitochondrial membrane potential (MMP) in mammalian cells. With JC-10 dye and fluorescence-based detection, the kit enables precise monitoring of apoptosis and the effects of apoptosis inhibitors or activators. Whether for basic research, drug screening, or pharmacological studies, this kit offers a robust solution for studying mitochondrial function and cell viability.