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Cat. No.: MDRRFM-500 (for 50-500T)

Cat. No.: MDRRFM-2k (for 200-2000T)

Description

Mitochondrial Deep Red Fluorescence Staining Kit, utilizing the deep red fluorescent probe Mito-Tracker Deep Red FM, is primarily used for the specific fluorescent staining of mitochondria in live cells. This kit is suitable for use with fluorescence microscopy, laser confocal microscopy, and flow cytometry. The kit cannot be used for staining fixed cells, but cells can be fixed after staining.

Mito-Tracker Deep Red FM is a cell-permeable, deep red fluorescent probe that specifically labels biologically active mitochondria and detects mitochondrial membrane potential. This high-purity probe is compatible with subsequent cell fixation and permeabilization, though fluorescence intensity may decrease after these processes.

As a deep 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 2-8 fold). The excitation and emission spectra of this probe do not overlap with those of common green and red fluorescent probes, making it ideal for multicolor fluorescence staining experiments.

Mito-Tracker Deep Red FM can be used as a mitochondria-specific fluorescent probe. Similar to Rhodamine 123 or JC-1, Mito-Tracker Deep Red FM's staining of mitochondria depends on mitochondrial membrane potential. Therefore, the probe can only stain live cells or tissues and cannot stain fixed or permeabilized cells or tissues. As Mito-Tracker Deep Red FM aggregates within 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.

Mito-Tracker Deep Red FM has a molecular formula of C34H36Cl2N2 and a molecular weight of 543.58. It is a deep red fluorescent probe with a maximum excitation wavelength of 644 nm and a maximum emission wavelength of 665 nm.

Components

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

Storage

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

Precautions

• Repeated freeze-thaw cycles of Mito-Tracker Deep Red FM (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 Deep Red FM.
• 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.