MitoSOX Red |
| Katalog-Nr.GC68230 |
MitoSOX Red is a live cell fluorescent probe targeting mitochondria with maximum excitation/emission light of 510/580 nm.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 1003197-00-9
Sample solution is provided at 25 µL, 10mM.
MitoSOX Red is a live cell fluorescent probe targeting mitochondria with a maximum excitation/emission light of 510/580 nm. MitoSOX Red has cell membrane permeability and can quickly and selectively target mitochondria after entering cells. After entering mitochondria, MitoSOX Red is easily oxidized by superoxide and produces strong red fluorescence after combining with nucleic acids in mitochondria. MitoSOX Red can be used as a fluorescent indicator to specifically detect superoxide[1-2].
Dihydroethidium, also called hydroethidine (HE), can be oxidized by reactive species and subsequently binds to DNA to produce fluorescence. MitoSOX Red is a dihydroethidium derivative with a cationic triphenylphosphonium moiety. This positively charged probe rapidly accumulates in mitochondria and can therefore be used to monitor superoxide/ROS production within mitochondria by fluorometry, microscopy, or flow cytometry[3]. For example, in NK cells, MitoSOX red can observe the distribution of mitochondrial ROS[1]. Confocal microscopy imaging shows that in H9c2 cells treated with paraquat, the fluorescence intensity of MitoSOX Red-labeled mitochondria increases significantly[4].
References:
[1] Jin F, Wu Z, Hu X, Zhang J, Gao Z, Han X, Qin J, Li C, Wang Y. The PI3K/Akt/GSK-3β/ROS/eIF2B pathway promotes breast cancer growth and metastasis via suppression of NK cell cytotoxicity and tumor cell susceptibility. Cancer Biol Med. 2019 Feb;16(1):38-54.
[2] Milliken A S, Nadtochiy S M, Brookes P S. Inhibiting succinate release worsens cardiac reperfusion injury by enhancing mitochondrial reactive oxygen species generation[J]. Journal of the American Heart Association, 2022, 11(13): e026135.
[3] Kauffman ME, Kauffman MK, Traore K, Zhu H, Trush MA, Jia Z, Li YR. MitoSOX-Based Flow Cytometry for Detecting Mitochondrial ROS. React Oxyg Species (Apex). 2016;2(5):361-370.
[4] Mukhopadhyay P, Rajesh M, Yoshihiro K, et al. Simple quantitative detection of mitochondrial superoxide production in live cells[J]. Biochemical and biophysical research communications, 2007, 358(1): 203-208.
This plan only provides a guide, please modify it to meet your specific needs.
1. Prepare MitoSOX Red staining solution
(1) This product is provided in the form of a 5mM DMSO stock solution. It is recommended to centrifuge the product before first use, fill the tube with nitrogen after aliquots, and store it in the dark at -20°C or -80°C.
(2) Dye working solution: Dilute the stock solution with a suitable buffer (such as serum-free medium or PBS) to prepare a MitoSOX Red working solution with a concentration of 1-10μM.
Note: Please adjust and optimize the working fluid concentration according to the actual situation, and prepare it now.
2. Cell suspension staining (taking 6-well plate as an example)
(1) Centrifuge suspended cells at 1000g for 3-5 minutes. Discard the supernatant and wash twice with PBS for 5 minutes each time.
(2) Wash the adherent cells twice with PBS, add trypsin to digest the cells, and centrifuge at 1000g for 3-5 minutes after digestion is completed.
(3) Add 1 mL of dye working solution to resuspend the cells, and incubate at room temperature in the dark for 5-30 minutes. The optimal culture time for different cells is different.
(4) After the incubation, centrifuge at 1000g for 5 minutes, remove the supernatant, and add PBS to wash 2-3 times, 5 minutes each time.
(5) Use serum-free cell culture medium or PBS to resuspend the cells and observe them through fluorescence microscopy or flow cytometry.
3. Cell adhesion staining+
(1) Culture adherent cells on sterile coverslips.
(2) Remove the coverslip from the culture medium, aspirate the excess culture medium, and place the coverslip in a humid environment.
(3) Add 100μL dye working solution from one corner of the coverslip, shake gently to evenly cover all cells with the dye, and incubate at room temperature in the dark for 5-30 minutes.
(4) Aspirate away the dye working solution and use culture solution to wash the coverslip 2 to 3 times for 5 minutes each time.
4. Microscope detection: The maximum excitation/emission light of MitoSOX Red is 396/610nm. MitoSOX Red dye can also excite at 510nm. If MitoSOX Red dye excites at 396nm instead of 510nm, it may be that the dye is more selective for labeling mitochondrial superoxide.
Precautions:
(1) Fluorescent dyes all have quenching problems. Please avoid light as much as possible to slow down fluorescence quenching.
(2) For your safety and health, please wear a lab coat and disposable gloves.
| Cas No. | 1003197-00-9 | SDF | |
| Formula | C43H43IN3P | M.Wt | 759.7 |
| Löslichkeit | DMSO : 150 mg/mL (197.45 mM; Need ultrasonic) | Storage | Store at -20°C,protect from light,stored under nitrogen |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
|
1 mg | 5 mg | 10 mg |
| 1 mM | 1.3163 mL | 6.5815 mL | 13.1631 mL |
| 5 mM | 0.2633 mL | 1.3163 mL | 2.6326 mL |
| 10 mM | 0.1316 mL | 0.6582 mL | 1.3163 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 30 reference(s) in Google Scholar.)
GLPBIO products are for RESEARCH USE ONLY. Please make sure your review or question is research based.
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