ROS fluorescent probe DHE |
Catalog No.GD20803
One-Click Copy Product Info
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ROS fluorescent probe DHE is the commonly used fluorescent dyes to measure ROS in cells and tissues at 480nm wavelength excitation and 610nm wavelength emission.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 38483-26-0
Sample solution is provided at 25 µL, 10mM.
ROS fluorescent probe DHE is the commonly used fluorescent dyes to measure ROS in cells and tissues at 480nm wavelength excitation and 610nm wavelength emission [1]. ROS fluorescent probe DHE can be rapidly absorbed by cells. After specifically reacting with superoxide free radicals, it will generate a red fluorescent product, 2-hydroxyethidium[2]. ROS fluorescent probe DHE has been widely used in the development of novel high-performance liquid chromatography (HPLC) methods for the rapid detection of superoxide in intact cells and tissues[3].
References:
[1] Kumar R, Gullapalli R R. High throughput screening assessment of reactive oxygen species (ROS) generation using Dihydroethidium (DHE) fluorescence dye[J]. Journal of visualized experiments: JoVE, 2024 (203): 10.3791/66238.
[2] Zhu L, Lu M, Yin X. Ultrasensitive determination of intracellular superoxide in individual HepG2 cells by microfluidic chip electrophoresis[J]. Talanta, 2008, 75(5): 1227-1233.
[3] Fink B, Laude K, McCann L, et al. Detection of intracellular superoxide formation in endothelial cells and intact tissues using dihydroethidium and an HPLC-based assay[J]. American Journal of Physiology-Cell Physiology, 2004, 287(4): C895-C902.
Evaluation of reactive oxygen species (ROS) generation using ROS fluorescent probe DHE
This protocol only provides a guideline, and should be modified according to your specific needs
1. Stock and dilute solution preparation for DHE staining of cells
Dilute the DHE stock solution (5mM in DMSO) with PBS or normal saline to a working solution with a final concentration of 10μM. NOTE: The working solution must be prepared fresh on the day of the experiment.
2. Oxidative stress treatment in cells
HepG2 cells reaching 80% to 90% confluence were treated with H2O2 (0, 250μM, 500μM, 750μM, and 1000μM) and Menadione (0, 25μM, 50μM, 75μM, and 100μM) for 30 Minutes to induce representative oxidative stress in 96-well plates.
3. DHE staining process
(1) Remove the media from wells and wash gently once with either 1×PBS or DMEM.
(2) Add 100μl of DMEM media containing 10μM DHE (working solution) into each well and incubate at 37°C for 30min.
(3) After 30min incubation period, remove the DHE-containing media and wash each well gently three times with the 1×PBS.
(4) Add 200μl of 1×PBS to each well with Hoechst 33342 nuclear staining dye for 10min at room temperature.
(5) Remove the Hoechst 33342 nuclear staining solution from the well and add 200μl of 1×PBS to each well.
(6) Move the plate over to the high-content screening platform, fluorescence microscopy, or plate reader for image acquisition.
4. Experimental Precautions
(1) DHE should be stored in the dark throughout the experiment.
(2) This protocol and all reagents are for scientific research use only.
References:
[1] Kumar R, Gullapalli R R. High throughput screening assessment of reactive oxygen species (ROS) generation using Dihydroethidium (DHE) fluorescence dye[J]. Journal of visualized experiments: JoVE, 2024 (203): 10.3791/66238.
References:
[1] Kumar R, Gullapalli R R. High throughput screening assessment of reactive oxygen species (ROS) generation using Dihydroethidium (DHE) fluorescence dye[J]. Journal of visualized experiments: JoVE, 2024 (203): 10.3791/66238.
| Cas No. | 38483-26-0 | SDF | |
| Formula | C21H21N3 | M.Wt | 315 |
| Solubility | 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 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 3.1746 mL | 15.873 mL | 31.746 mL |
| 5 mM | 634.9 μL | 3.1746 mL | 6.3492 mL |
| 10 mM | 317.5 μL | 1.5873 mL | 3.1746 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 saline, 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% Appearance: A solution
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 30 reference(s) in Google Scholar.)