Resorufin

Resorufin (NSC 12097) is a highly fluorescent pink dye. Resorufin shows favorable properties such as water solubility, red-shifted absorption/emission signals. Resorufin is used for the detection of ROS/RNS and a second analyte^[1][2]. In Vitro:Preparation of Resorufin staining solution
1.1 Preparation of stock solution
Prepare 1000 μM Resorufin with DMSO.
Note: 1) Resorufin stock solution is recommended to be stored in aliquots at -20°C in the dark.
1.2 Preparation of working solution
Prepare 0.4 μM Resorufin working solution in PBS (pH 7.4).
Resorufin staining
1) Pre-treat HepG2 cells with CD for 30 min, then remove the culture medium. Thereafter, wash the cells three times with 200 μL PBS, then add 150 μL fresh DMEM and 15 μL 22 μM Resazurin solution (diluted in DMEM) to each well. After incubation at 37 °C in the dark for 30 min, measure the Resorufin content using a plate reader.
2) HepG2 cells were pretreated with CD for 120 min, and then the culture medium was removed. Thereafter, the cells were washed three times with 200 μL PBS, and then 150 μL fresh DMEM and 15 μL 22 μM resazurin solution (diluted in DMEM) were added to each well. After incubation at 37 °C in the dark for 120 min, the resorufin content was determined by a plate reader.
3) Data analysis: The Benesi-Hildebrand equation was used, where the fluorescence intensity values ??were replaced by absorbance values.