MitoDNA™ Red 610
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
There are very few probes that can be effectively used to detect mitochondrial deoxyribonucleic acid (mtDNA). The common fluorescent DNA probes (such as DAPI, Hoechst or SYBR® Green) lack the specificity to target mitochondria. They predominantly stain nuclei. MitoDNA™ Red 610 is a cell permeable dye that specifically stains mtDNA in live cells. It provides an efficient way of labeling mtDNA for the dynamic imaging of mtDNA in live cells. MitoDNA™ Red 610 has a large Stokes Shift that gives great signal-to-noise ratio and can be easily used for multiplex staining in cells with other fluorescent imaging probes. mtDNA is a small circular DNA found within mitochondria present in the cytoplasm of a cell. This DNA is supplementary to the nucleic acid material found in the nucleus of each cell. The mtDNA codes for 37 genes that promote the proper functioning of some cells. The mitochondria synthesize adenosine triphosphate (ATP) through oxidative phosphorylation and encode information for the synthesis of enzymes, transfer ribonucleic acid (tRNA), and ribosomal RNA (rRNA). Disorders of mtDNA and mutations in its genes can predispose to health problems like age-related hearing loss, diabetes, and brain, heart, and liver failure, among other conditions. Moreover, mtDNA and its associated mitochondrial disorders can predispose people to different types of cancers including lymphomas, leukemias, and breast, intestine, liver and kidney tumors etc.
Example protocol
AT A GLANCE
Before using MitoDNA™ Red 610 for the first time, allow it to thaw at room temperature. Then, briefly centrifuge it to collect the dried pellet.
Prepare cells in a growth medium.
Stain cells with MitoDNA™ Red 610 working solution.
Incubate samples for 5 to 15 minutes at 37 °C.
Monitor fluorescence intensity at Ex/Em = 490/610 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
Prepare a 5 to 10 mM MitoDNA™ Red 610 stock solution in DMSO. For example, add 290 μL of DMSO to the MitoDNA™ Red 610 vial to create a 10 mM stock solution.
Note: Prepare a single aliquot of the unused MitoDNA™ Red 610 stock solution and store it at ≤ -20 º C, protected from light. Avoid repeated freeze-thaw cycles.
PREPARATION OF WORKING SOLUTION
Prepare a 5 to 10 μM working solution by diluting the MitoDNA™ Red 610 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
Plate the cells in a 96-well plate with black walls and a clear bottom.
Remove the cell culture medium and add 100 µL of MitoDNA™ Red 610 working solution directly to the cells.
Incubate the cells at 37°C for 5-15 minutes, protected from light.
Note: The concentration and incubation time of MitoDNA™ Red 610 may vary depending on the cell line. Test different concentrations to determine the optimal dose.
Remove the dye working solution and wash the cells twice with HHBS buffer.
Add HHBS buffer and analyze the cells using a fluorescence microscope with excitation/emission settings of 490/610 nm.
Spectrum
Product family
Name | Excitation (nm) | Emission (nm) |
MitoDNA™ Red 680 | 597 | 681 |
MitoDNA™ Red 710 | 511 | 707 |
References
Authors: Bao, Yu and Yang, Shiyuan and Zhao, Hailan and Wang, Yezhen and Li, Ke and Liu, Xue and Zhang, Wei and Zhu, Xue
Journal: BMC pulmonary medicine (2024): 176
Authors: Zhang, Yingchi and Xu, Tao and Xie, Jie and Wu, Hua and Hu, Weihua and Yuan, Xuefeng
Journal: Cell communication and signaling : CCS (2024): 240
Authors: Dancis, Andrew and Pandey, Ashutosh K and Pain, Debkumar
Journal: Biochimica et biophysica acta. Molecular cell research (2024): 119733
Authors: Lei, Tang and Rui, Yu and Xiaoshuang, Zhou and Jinglan, Zhang and Jihong, Zhang
Journal: Cell death discovery (2024): 168
Authors: Feng, Fuhai and He, Shasha and Li, Xiaoling and He, Jiake and Luo, Lianxiang
Journal: Aging and disease (2024): 714-738