Calculator
MitoDNA™ Green 530
Product key features
- mtDNA-Specific: Selectively binds to mtDNA without nuclear interference
- Live-Cell Compatibility: Easily permeates cells for real-time mtDNA tracking
- High Signal Clarity: Large Stokes shift ensures a strong signal-to-noise ratio for multiplex imaging
- Broad Research Applicability: Ideal for investigating mtDNA-linked diseases and mitochondrial function
Product description
Detecting mitochondrial DNA (mtDNA) has been challenging due to the non-specific nature of conventional DNA probes like DAPI, Hoechst, or SYBR® Green, which primarily stain the nucleus. MitoDNA™ Green 530 overcomes this limitation with its high specificity for mtDNA, allowing researchers to selectively visualize and track mitochondrial dynamics in live cells. This cell-permeable dye offers a large Stokes Shift, providing a high signal-to-noise ratio ideal for multiplex imaging alongside other fluorescent probes.
mtDNA is a small, circular DNA found in the mitochondria of a cell, separate from the nuclear DNA. It encodes 37 genes essential for mitochondrial function, including those involved in ATP synthesis, enzyme production, and tRNA and rRNA synthesis. Mutations or disorders in mtDNA can lead to various health issues, including age-related hearing loss, diabetes, and failures of the brain, heart, and liver. Additionally, mtDNA mutations are linked to an increased risk of cancers, such as lymphomas, leukemias, and tumors in the breast, intestine, liver, and kidneys
Example protocol
AT A GLANCE
Before initial use, thaw MitoDNA™ Green 530 at room temperature and briefly centrifuge to collect the dried pellet.
Prepare cells in growth medium.
Stain cells with MitoDNA™ Green 530 working solution.
Incubate samples at 37°C for 5–15 minutes.
Monitor fluorescence intensity with FITC filter.
PREPARATION OF STOCK SOLUTIONS
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles
Prepare a 5–10 mM MitoDNA™ Green 530 stock solution in DMSO. For example, add 236 μL of DMSO to the MitoDNA™ Green 530 vial to create a 10 mM stock solution.
Note: Prepare single-use aliquots of the stock solution and store at ≤ -20°C. Protect from light and avoid repeated freeze-thaw cycles.
PREPARATION OF WORKING SOLUTION
Prepare a 5–10 μM working solution by diluting the MitoDNA™ Green 530 stock solution in Hanks solution with 20 mM Hepes buffer (HHBS, AAT cat# 20011).
Note: For optimal results, use this solution within a few hours of preparation.
Note: Protect the working solution from light by covering it with foil or placing it in the dark.
SAMPLE EXPERIMENTAL PROTOCOL
Plate cells as needed in a 96-well black wall, clear bottom plate.
Remove the cell culture medium and add 100 µL of MitoDNA™ Green 530 working solution to each well.
Incubate cells at 37°C in a 5% CO2 incubator for 5–15 minutes, keeping them protected from light.
Note: The optimal concentration and incubation time may vary by cell line; we recommend testing different concentrations.
Remove the dye working solution and wash the cells twice with HHBS buffer.
Add HHBS buffer and analyze cells using a fluorescence microscope with a FITC filter set.
Spectrum
References
Authors: Li, Hu and Shangqing, Zhang and Yae, Zhao and Fan, Yang and Xinyue, Zhang and Shirui, Liu and Tianyi, Zhang and Dongling, Niu
Journal: Gene (2025): 148981
Authors: Feiz-Haddad, Mohammad Hossein and Moradkhani, Mohammad-Ali
Journal: Comparative immunology, microbiology and infectious diseases (2024): 102210
Authors: Albrechtová, Martina and Kašparová, Eva Štefková and Langrová, Iva and Hart, Vlastimil and Neuhaus, Birger and Jankovská, Ivana and Petrtýl, Miroslav and Magdálek, Jan and Špakulová, Marta
Journal: Frontiers in veterinary science (2024): 1388292
Authors: Hidalgo-Gutierrez, Agustin and Shintaku, Jonathan and Ramon, Javier and Barriocanal-Casado, Eliana and Pesini, Alba and Saneto, Russell P and Garrabou, Gloria and Milisenda, Jose Cesar and Matas-Garcia, Ana and Gort, Laura and Ugarteburu, Olatz and Gu, Yue and Koganti, Lahari and Wang, Tian and Tadesse, Saba and Meneri, Megi and Sciacco, Monica and Wang, Shuang and Tanji, Kurenai and Horwitz, Marshall S and Dorschner, Michael O and Mansukhani, Mahesh and Comi, Giacomo Pietro and Ronchi, Dario and Marti, Ramon and Ribes, Antonia and Tort, Frederic and Hirano, Michio
Journal: Annals of neurology (2024)
Authors: Valsecchi, Elena
Journal: Molecular ecology resources (2024): e14012
Safety data sheet