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Dihydrorhodamine 123 (DHR 123) (Synonyms: DHR 123)

カタログ番号GC30581

ジヒドロローダミン 123 (DHR 123) (DHR 123) は、蛍光プローブ (Λex=488 nm、Λem=525 nm) です。

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Dihydrorhodamine 123 (DHR 123) 化学構造

Cas No.: 109244-58-8

サイズ 価格 在庫数 個数
1mg
$45.00
在庫あり
5mg
$108.00
在庫あり

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Sample solution is provided at 25 µL, 10mM.

Product Documents

Quality Control & SDS

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Protocol

This plan only provides a guide, please modify it to meet your specific needs.

1.Solution Preparation:

(1) Preparation of stock solution: Allow the solid stored at low temperature to equilibrate to room temperature. Dissolve Dihydrorhodamine 123 in DMSO to prepare a stock solution with a concentration of 5 mM.

Note: Unused stock solutions should be aliquoted and stored in the dark at -20°C or -80°C to avoid repeated freeze-thaw cycles.

 

(2) Preparation of working solution: Before the formal experiment, dilute the stock solution to the desired working concentration using an appropriate buffer (e.g., serum-free medium or PBS).

Note: The optimal working concentration should be adjusted based on the actual situation or determined by consulting the literature and setting up a gradient concentration experiment. The working solution must be prepared fresh and used immediately.

 

Using DHR123 to Detect Intracellular ROS Steps [1] (from literature, for reference only)

(1) Cell treatment: Wash the cells with water to remove any contaminants or debris.

(2) DHR123 incubation: Resuspend the cells in PBS containing DHR123 (15 μg/ml, approximately 43μM). Incubate with shaking at 30°C for 90 minutes.

(3) Washing cells: Wash the cell pellet twice with PBS buffer to remove excess DHR123.

(4) Fluorescence detection: Measure the fluorescence of 10,000 cells using a flow cytometer. Set the excitation and emission wavelengths to 485 nm and 535 nm, respectively, to detect the fluorescence emitted by the reaction of DHR123 with intracellular ROS. Analyze the fluorescence distribution in the samples using FlowJo 7.6.5 software.

Using DHR123 to Detect Intracellular ROS Steps [2] (from literature, for reference only)

(1) Cell treatment: Treat cells according to experimental requirements.

(2) DHR123 incubation: Prepare a solution containing 2.5 μM DHR123 in HBSS buffer solution. Incubate the cells with DHR123 solution at 37°C for 30 minutes.

(3) Washing cells: Wash the cells twice with HBSS to remove excess DHR123.

(4) Lysing cells: Lyse the cells with RIPA buffer to release the intracellularly converted Rh123+.

(5) Fluorescence detection: Set the excitation and emission wavelengths to 485 nm and 535 nm, respectively, to detect Rh123+ fluorescence. Record the fluorescence intensity readings as relative units indicating intracellular ROS levels.

Precautions: This experimental protocol provides guidance for detecting intracellular reactive oxygen species (ROS) using Dihydrorhodamine 123 (DHR123) and subsequent fluorescence measurement. Adjustments can be made based on specific experimental requirements.

References:

[1]Antoine Simoneau, Étienne Ricard, Sandra Weber, et al. Chromosome-wide histone deacetylation by sirtuins prevents hyperactivation of DNA damage-induced signaling upon replicative stress [J] Nucleic Acids Research, Volume 44, Issue 6, 7 April 2016, Pages 2706–2726.

[2] 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.

Background

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 485 nm, and its emission wavelength is 535 nm[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).

Chemical Properties

Cas No. 109244-58-8 SDF
同義語 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
溶解度 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.
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 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
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**ストックソリューションを準備する際には、常にバイアルラベルおよび MSDS/CoA(オンラインで利用可能)で掲載された製品のロット固有分子量を使用してください。

計算

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

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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.)

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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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Review for Dihydrorhodamine 123 (DHR 123)

Average Rating: 5 ★★★★★ (Based on Reviews and 38 reference(s) in Google Scholar.)

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