Dihydrorhodamine 123 (DHR 123) (Synonyms: DHR 123) |
| Catalog No.GC30581 |
La dihidrorrodamina 123 (DHR 123) (DHR 123) es una sonda fluorescente (Λex=488 nm, Λem=525 nm).
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 109244-58-8
Sample solution is provided at 25 µL, 10mM.
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Dihydrorhodamine 123 (DHR 123) is a non-fluorescent precursor that transforms into the fluorescent compound rhodamine 123 (Rh123+) upon oxidation. This transformation process makes DHR 123 a useful tool for detecting reactive oxygen species (ROS) within cells. Dihydrorhodamine 123 can cross the cell membrane, and once inside the cell, it is oxidized by ROS into Rh123+, producing green fluorescence that accumulates in the mitochondrial matrix. The excitation wavelength of Rh123+ is 485nm, and its emission wavelength is 528nm[1]. Dihydrorhodamine 123 can also be used as an indicator for the formation of intracellular peroxynitrite[2].
References:
[1] Wing-Kee Lee, Stephanie Probst, Bettina Scharner, et al. Distinct concentration-dependent oxidative stress profiles by cadmium in a rat kidney proximal tubule cell line[J] Archives of Toxicology. 30 January 2024.
[2] Crow, J.P. Dichlorodihydrofluorescein and dihydrorhodamine 123 are sensitive indicators of peroxynitrite in vitro: Implications for intracellular measurement of reactive nitrogen and oxygen species Nitric Oxide1(2)145-157(1997).
This plan only provides a guide, please modify it to meet your specific needs.
1. Preparation of staining solution
(1) Prepare stock solution: Dissolve Dihydrorhodamine 123 (DHR 123) in DMSO and prepare a stock solution with a concentration of 1-10mM.
Note: Unused stock solution should be aliquoted and stored in the dark at -20°C or -80°C to avoid repeated freezing and thawing.
(2) Prepare working solution: Dilute the stock solution with a suitable buffer (such as serum-free medium or PBS) and prepare a working solution with a concentration of 1-10μM.
Note: Please adjust the concentration of the working fluid according to the actual situation and prepare it now.
2. Cell suspension staining
(1) Suspension cells: Centrifuge at 1000g for 3-5 minutes at 4°C, discard the supernatant, and wash twice with PBS for 5 minutes each time.
(2) Adherent cells: Wash twice with PBS, add trypsin to digest the cells, and centrifuge at 1000g for 3-5 minutes after digestion is completed.
(3) Add DHR 123 working solution to resuspend the cells, and incubate at room temperature in the dark for 5-30 minutes. The optimal incubation time for different cells is different, please explore by yourself according to the specific experimental needs.
(4) After the incubation, centrifuge at 1000g for 5 minutes, remove the supernatant, add PBS and wash 2-3 times, 5 minutes each time.
(5) Resuspend cells in pre-warmed serum-free cell culture medium or PBS and observe by fluorescence microscope or flow cytometry.
3. Cell adhesion staining
(1) Culture adherent cells on sterile coverslips.
(2) Remove the coverslip from the culture medium, suck out 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 and shake gently to evenly cover all cells with the dye.
(4) Incubate at room temperature in the dark for 5-30 minutes. The optimal incubation time for different cells is different, please explore by yourself according to the specific experimental needs.
(5) After the incubation, discard the dye working solution and use pre-warmed culture solution to wash the coverslip 2 to 3 times.
4. Microscope detection: The maximum excitation/emission wavelength of DHR 123 is 485/528nm.
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. | 109244-58-8 | SDF | |
| Sinónimos | DHR 123 | ||
| Canonical SMILES | O=C(OC)C1=CC=CC=C1C2C3=C(OC4=C2C=CC(N)=C4)C=C(N)C=C3 | ||
| Formula | C21H18N2O3 | M.Wt | 346.38 |
| Solubility | DMSO: 100 mg/mL (288.70 mM) | Storage | Store at -20°C,protect from light |
| 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 | 2.887 mL | 14.435 mL | 28.87 mL |
| 5 mM | 0.5774 mL | 2.887 mL | 5.774 mL |
| 10 mM | 0.2887 mL | 1.4435 mL | 2.887 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 38 reference(s) in Google Scholar.)
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