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Cat. No.: MDRCMX-500 (for 50-500T)

Cat. No.: MDRCMX-2k (for 200-2000T)

Description

Mitochondrial Red Fluorescence Staining Kit, also known as the Mitochondrial Membrane Potential Assay Kit with Mito-Tracker Red CMXRos, utilizes the red fluorescent probe Mito-Tracker Red CMXRos for the specific fluorescent staining of mitochondria in live cells or the detection of mitochondrial membrane potential. This kit is suitable for use with fluorescence microscopy, laser confocal microscopy, and flow cytometry. The kit is compatible with subsequent cell fixation and permeabilization, though fluorescence intensity of Mito-Tracker Red CMXRos may decrease after these processes.

Mito-Tracker Red CMXRos, also known as MitoTracker Red CMXRos, is a cell-permeable X-rosamine derivative (Chloromethyl-X-rosamine, abbreviated as CMXRos) that specifically labels biologically active mitochondria and detects mitochondrial membrane potential. This high-purity, high-quality probe is compatible with subsequent cell fixation and permeabilization, though fluorescence intensity may decrease after these processes.

As an oxidized red fluorescent dye, it easily incubates with cells, crossing cell membranes via passive transport, and specifically labels biologically active mitochondria through its mildly thiol-reactive chloromethyl functional group. This group reacts covalently with thiol groups on mitochondrial proteins, ensuring that mitochondrial fluorescence labeling persists even after treatment with aldehyde fixatives like paraformaldehyde or formaldehyde, and permeabilization agents like Triton X-100, albeit with some fluorescence reduction (approximately 2-10 fold). Due to minimal overlap between the excitation and emission spectra of this probe and common green fluorescent probes, it is ideal for dual-labeling experiments.

Mito-Tracker Red CMXRos has a molecular formula of C32H32Cl2N2O, a molecular weight of 531.5, and a CAS number of 167095-09-2. It emits red fluorescence with a maximum excitation wavelength of 579 nm and a maximum emission wavelength of 599 nm. Mito-Tracker Red CMXRos can be used as a mitochondria-specific fluorescent probe. Similar to Rhodamine 123 or JC-1, Mito-Tracker Red CMXRos's staining of mitochondria depends on mitochondrial membrane potential. Therefore, this probe can only stain live cells or tissues and cannot stain fixed or permeabilized cells or tissues. As Mito-Tracker Red CMXRos accumulates in mitochondria depending on mitochondrial membrane potential, it can also serve as an indicator probe for mitochondrial membrane potential and be used to detect apoptosis by monitoring changes in mitochondrial membrane potential.

For samples in a 96-well plate, using 100 µl of staining solution per well, the small and medium packaging of this kit can perform 500 and 2000 tests, respectively. For flow cytometry, assuming a sample volume of 0.5 ml per test, the small and medium packaging can perform 100 and 400 tests, respectively. For adherent cells in a 6-well plate, using 1 ml of staining solution per well, the small and medium packaging can perform 50 and 200 tests, respectively.

Components

• Mito-Tracker Red CMXRos (1000X)
• Hoechst 33342 (1000X)
• FCCP (1000X)
• Assay Buffer

Storage

Store at -20ºC for one year. Mito-Tracker Red CMXRos (1000X) and Hoechst 33342 (1000X) must be protected from light.

Precautions

• Repeated freeze-thaw cycles of Mito-Tracker Red CMXRos (1000X) can affect its performance. Please aliquot appropriately to avoid repeated freeze-thaw cycles.
• It is recommended to detect the staining as soon as possible after staining. Prolonged periods may increase the number of apoptotic or necrotic cells and significantly alter mitochondrial morphology.
• This kit is suitable for mitochondrial fluorescence labeling in live cells but is not suitable for labeling fixed cells. For post-fixation labeling, we recommend Mitochondrial Green Fluorescence Staining Kit with Mito-Tracker Green. After fixation, fluorescence intensity may decrease, so it is advisable to increase the concentration of Mito-Tracker Red CMXRos.
• For small volumes of liquid, centrifuge for a few seconds before each use to ensure the liquid settles to the bottom of the tube.
• All fluorescent dyes are subject to quenching, so protect from light as much as possible to reduce fluorescence quenching.
• This product is for scientific research use by professionals only and is not intended for clinical diagnosis or treatment, food, or pharmaceutical use. Do not store in ordinary residences.
• For your safety and health, please wear a lab coat and disposable gloves when handling.