Mito-Tracker Green |
Catalog No.GC26229 |
Mito-Tracker Green is a green fluorescent probe for mitochondria in living cells with a maximum excitation/emission wavelength of 490/516 nm.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 201860-17-5
Sample solution is provided at 25 µL, 10mM.
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Protocol
This plan only provides a guide, please modify it to meet your specific needs.
1. Prepare dyeing solution
(1) Dye stock solution: Use DMSO to dissolve Mito-Tracker Green into a 1-5mM stock solution. The prepared stock solution is aliquoted and stored in the dark at -20°C.
(2) Dye working solution: Dilute the stock solution with a suitable buffer (such as serum-free medium, HBSS or PBS) to prepare an Mito-Tracker Green working solution with a concentration of 20-200 nM.
Attention:
① It is recommended to use a relatively low concentration for staining. If a higher concentration is used, it may stain other cell structures. To reduce false positives caused by excessive probe loading, it is recommended to use a low concentration as much as possible without affecting the staining effect;
② If cells are incubated in a dye free medium after staining, attenuation of fluorescence signals and vacuolization of cells will be observed;
③ Please adjust and optimize the concentration of the working solution according to the actual situation, and prepare it as needed.
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 15-45minutes. 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 100 μL of 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 15-45 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. Observe using a fluorescence microscope or flow cytometer. Mito-Tracker Green has a maximum excitation/emission wavelength of 490/516nm.
Precautions:
① Mito-Tracker Green is only used to stain living cells, and fixation will cause the fluorescence to disappear;
② Mito-Tracker Green working fluid needs to be pre-warmed at 37°C before use;
③ If the dyeing effect is not good, it is recommended to increase the concentration of the working solution or appropriately extend the dyeing time within the recommended time range;
④ Fluorescent dyes all have quenching problems. Please try to avoid light to slow down fluorescence quenching;
⑤ For your safety and health, please wear a lab coat and disposable gloves.
Mito-Tracker Green is a mitochondrial green fluorescent probe that can be used for mitochondria-specific fluorescent staining of living cells. After entering cells, Mito-Tracker Green selectively accumulates in the mitochondrial matrix and covalently binds to mitochondrial proteins by reacting with free sulfhydryl groups of cysteine residues. The maximum excitation/emission wavelength of Mito-Tracker Green is 490/516 nm.
Cas No. | 201860-17-5 | SDF | |
Formula | C34H28C15N3O | M.Wt | 671.88 |
Solubility | Storage | Store at -20 ℃ away from light, effective for six months. | |
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. |
||
Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. |
Complete Stock Solution Preparation Table
Prepare stock solution | |||
1 mg | 5 mg | 10 mg | |
1 mM | 1.4884 mL | 7.4418 mL | 14.8836 mL |
5 mM | 0.2977 mL | 1.4884 mL | 2.9767 mL |
10 mM | 0.1488 mL | 0.7442 mL | 1.4884 mL |
In vivo Formulation Calculator (Clear solution)
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
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.
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