Dihydrorhodamine 123 (solution) 

Dihydrorhodamine 123 (DHR 123) (solution) is a non-fluorescent reactive oxygen species (ROS) indicator. Dihydrorhodamine 123 is oxidized to fluorescent Rhodamine 123 (HY-D0816) within cells in the presence of reactive oxygen species and it localizes in mitochondria (Ex/Em = 515/536 nm).
Solvent and concentration: DMSO: 10 mM

Guide (The following is our recommended solution. This solution is merely a guideline and should be modified according to your specific needs.)
1. Solution Preparation
1. Work Solution Preparation Dilute the cell culture medium to 1-10 μM (based on experimental optimization).
Note: The working solution should be prepared and used immediately, and stored in a dark place.

2. Cell Staining 2.1 Treat the cells with reactive oxygen inducers or inhibitors. Note: If cells are treated with an oxidizing agent, rinse the cells three times with buffer solution or culture medium before staining to remove the compound and prevent the dye from being directly oxidized by the compound.
2.2 Dilute the Dihydrorhodamine 123 dye solution in serum-free medium to a final concentration of 5 μM.
2.3 Remove the cell culture medium and replace it with a medium containing the dye.
2.4 Incubate at 37°C for 30 minutes.
2.5 Detection can be performed using fluorescence microscopy or flow cytometry. No washing is required, but washing can be carried out to reduce the background in the microscopic examination.

When 10 μM Dihydrorhodamine 123 is present, by adding 50 nM phorbol 12-myristate 13-acetat (PMA) to stimulate the NADPH oxidase of neutrophils, the rate of rhodamine production can be increased. In the presence of 10 μM Dihydrorhodamine 123, the fluorescence intensity of the cells increases over time after adding 50 nM PMA. In the presence of 10 μM Dihydrorhodamine 123, the fluorescence of HL60 cells induced by adding 50 nM PMA continues to increase ^[1].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

346.39

C₂₁H₁₈N₂O₃

109244-58-8

Liquid (Density: 1.313±0.06 g/cm³)

Off-white to pink

O=C(OC)C1=CC=CC=C1C2C3=C(OC4=C2C=CC(N)=C4)C=C(N)C=C3

Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.

• [1]. Lydia M. Henderson et al. Dihydrorhodamine 123: a fluorescent probe for superoxide generation? Eur.J.Biochem. 217, 973-980.  [Content Brief]