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MitoDNA™ Red 710

The fluorescence response of MitoDNA™ Red 710 (5 µM) was assessed in HeLa cells before and after DNase treatment (2 units/reaction) at 37°C for 1 hour. Fluorescence intensities were measured using a fluorescence microscope equipped with a Violet long-pass filter (450 nm emission long-pass filter).
The fluorescence response of MitoDNA™ Red 710 (5 µM) was assessed in HeLa cells before and after DNase treatment (2 units/reaction) at 37°C for 1 hour. Fluorescence intensities were measured using a fluorescence microscope equipped with a Violet long-pass filter (450 nm emission long-pass filter).
The fluorescence response of MitoDNA™ Red 710 (5 µM) was assessed in HeLa cells before and after DNase treatment (2 units/reaction) at 37°C for 1 hour. Fluorescence intensities were measured using a fluorescence microscope equipped with a Violet long-pass filter (450 nm emission long-pass filter).
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Physical properties
Molecular weight342.91
SolventDMSO
Spectral properties
Excitation (nm)511
Emission (nm)707
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure

OverviewpdfSDSpdfProtocol


See also: Mitochondria
Molecular weight
342.91
Excitation (nm)
511
Emission (nm)
707
There are limited probes available that effectively detect mitochondrial DNA (mtDNA). Conventional fluorescent DNA probes, such as DAPI, Hoechst, or SYBR® Green, lack the specificity required for mitochondrial targeting and primarily stain nuclear DNA. MitoDNA™ Red 710 is a cell-permeable dye that selectively stains mtDNA in live cells, providing a method for dynamic imaging of mtDNA. This dye exhibits a large Stokes shift (Ex/Em = 510/710 nm), offering a high signal-to-noise ratio and enabling multiplex staining with other fluorescent probes. mtDNA is a small, circular DNA molecule located within the mitochondria in the cytoplasm. It supplements nuclear DNA and encodes 37 genes essential for mitochondrial and cellular functions. Mitochondria are responsible for ATP synthesis through oxidative phosphorylation and house the genetic information necessary for synthesizing key enzymes, transfer RNA (tRNA), and ribosomal RNA (rRNA). Mutations and disorders in mtDNA are implicated in a range of pathologies, including age-related hearing loss, diabetes, and organ dysfunctions in the brain, heart, and liver. Additionally, mtDNA mutations are associated with an elevated risk of various cancers, including lymphomas, leukemias, and tumors in the breast, intestines, liver, and kidneys.

Platform


Fluorescence microscope

Excitation510 nm
Emission710 nm
Recommended plateBlack wall/clear bottom

Example protocol


AT A GLANCE

Important Note

Before using MitoDNA™ Red 710 for the first time, allow it to thaw at room temperature. Then, briefly centrifuge it to collect the dried pellet.

Protocol Summary
  1. Prepare cells in a growth medium.

  2. Stain cells with MitoDNA™ Red 710 working solution.

  3. Incubate samples for 5 to 15 minutes at 37 °C.

  4. Monitor fluorescence intensity at Ex/Em = 510/710 nm.

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

MitoDNA™ Red 710 Stock Solution
  1. Prepare a 5 to 10 mM MitoDNA™ Red 710 stock solution in DMSO. For example, add 290 μL of DMSO to the MitoDNA™ Red 710 vial to create a 10 mM stock solution.

    Note: Prepare a single aliquot of the unused MitoDNA™ Red 710 stock solution and store it at ≤ -20 º C, protected from light. Avoid repeated freeze-thaw cycles.

PREPARATION OF WORKING SOLUTION

MitoDNA™ Red 710 Working Solution
  1. Prepare a 5 to 10 μM working solution by diluting the MitoDNA™ Red 710 stock solution in Hanks' solution with 20 mM HEPES buffer (HHBS).

    Note: For optimal results, use this solution within a few hours of preparation.

    Note: Cover the working solution with foil or store it in a dark place to protect it from light.

SAMPLE EXPERIMENTAL PROTOCOL

  1. Plate the cells in a 96-well plate with black walls and a clear bottom.

  2. Remove the cell culture medium and add 100 µL of MitoDNA™ Red 710 working solution directly to the cells.

  3. Incubate the cells at 37°C for 5-15 minutes, protected from light.

    Note: The concentration and incubation time of MitoDNA™ Red 710 may vary depending on the cell line. Test different concentrations to determine the optimal dose.

  4. Remove the dye working solution and wash the cells twice with HHBS buffer.

  5. Add HHBS buffer and analyze the cells using a fluorescence microscope with excitation/emission settings of 510/710 nm.

Calculators


Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of MitoDNA™ Red 710 to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

0.1 mg0.5 mg1 mg5 mg10 mg
1 mM291.622 µL1.458 mL2.916 mL14.581 mL29.162 mL
5 mM58.324 µL291.622 µL583.243 µL2.916 mL5.832 mL
10 mM29.162 µL145.811 µL291.622 µL1.458 mL2.916 mL

Molarity calculator

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Spectrum


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spectrum

Spectral properties

Excitation (nm)511
Emission (nm)707

Product Family


NameExcitation (nm)Emission (nm)
MitoDNA™ Red 610508607
MitoDNA™ Red 680597681

Images


References


View all 49 references: Citation Explorer
Release of damaged mitochondrial DNA: A novel factor in stimulating inflammatory response.
Authors: Li, Wenting and Li, Yuting and Zhao, Jie and Liao, Jiabao and Wen, Weibo and Chen, Yao and Cui, Huantian
Journal: Pathology, research and practice (2024): 155330
Mitochondrial DNA drives neuroinflammation through the cGAS-IFN signaling pathway in the spinal cord of neuropathic pain mice.
Authors: Huang, Penghui and Li, Li and Chen, Yaohua and Li, Yuping and Zhu, Dan and Cui, Jian
Journal: Open life sciences (2024): 20220872
High number of mitochondrial DNA alterations in postmortem brain tissue of patients with schizophrenia compared to healthy controls.
Authors: Bulduk, Bengisu K and Tortajada, Juan and Valiente-Pallejà, Alba and Callado, Luís F and Torrell, Helena and Vilella, Elisabet and Meana, J Javier and Muntané, Gerard and Martorell, Lourdes
Journal: Psychiatry research (2024): 115928
mtDNA-Server 2: advancing mitochondrial DNA analysis through highly parallelized data processing and interactive analytics.
Authors: Weissensteiner, Hansi and Forer, Lukas and Kronenberg, Florian and Schönherr, Sebastian
Journal: Nucleic acids research (2024)
Mitochondrial DNA copy number mediated the associations between perfluoroalkyl substances and breast cancer incidence: A prospective case-cohort study.
Authors: Feng, Yue and You, Yingqian and Li, Mengying and Guan, Xin and Fu, Ming and Wang, Chenming and Xiao, Yang and He, Meian and Guo, Huan
Journal: The Science of the total environment (2024): 173767
Mitochondrial DNA: Inherent Complexities Relevant to Genetic Analyses.
Authors: Ferreira, Tomas and Rodriguez, Santiago
Journal: Genes (2024)
Mitochondrial DNA Stress-Mediated Health Risk to Dibutyl Phthalate Contamination on Zebrafish (Danio rerio) at Early Life Stage.
Authors: Fan, Xiaoteng and Zhang, Dingfu and Hou, Tingting and Zhang, Qianqing and Tao, Lu and Bian, Chongqian and Wang, Zaizhao
Journal: Environmental science & technology (2024): 7731-7742
Exploring the Causal Effects of Mineral Metabolism Disorders on Telomere and Mitochondrial DNA: A Bidirectional Two-Sample Mendelian Randomization Analysis.
Authors: Feng, Zhijun and Wang, Yinghui and Fu, Zhengzheng and Liao, Jing and Liu, Hui and Zhou, Meijuan
Journal: Nutrients (2024)
Cells dispose of cytoplasmic mitochondrial DNA by nucleoid-phagy.
Authors:
Journal: Nature cell biology (2024)
Cytosolic Escape of Mitochondrial DNA Triggers cGAS-STING Pathway-Dependent Neuronal PANoptosis in Response to Intermittent Hypoxia.
Authors: Wang, Shuying and Tan, Jin and Zhang, Qiang
Journal: Neurochemical research (2024)