H2DCFDA is a cell-permeable probe used to detect intracellular reactive oxygen species (ROS).
ROS Measurements
For the detection of intracellular ROS level ,ROS-sensitive probe H2DCFDA is used. Adherent cells (ESCs, difESCs, eMSCs, HeLa, U118) are incubated with 5 µM staining solution in PBS in the dark for 30 min at 37°C, then harvested with 0.05% trypsin-EDTA solution, suspended in a fresh medium, and immediately analyzed with flow cytometer. Lymphocytes, both control and PHA-activated, are resuspended in PBS, incubated with 5 µM of H2DCFDA in the dark for 30 min at 37°C, and immediately analyzed. Along with the H2DCFDA probe, if indicated, ROS-insensitive modification of the fluorescent dye DCFDA is used. The staining procedure is the same as for the H2DCFDA.
Anat Rec (Hoboken). 2020 Jan 21.
Resibufogenin inhibited colorectal cancer cell growth and tumorigenesis through triggering ferroptosis and ROS production mediated by GPX4 inactivation
H2DCFDA purchased from AbMole
Species | Mouse | Rat | Rabbit | Guinea pig | Hamster | Dog |
Weight (kg) | 0.02 | 0.15 | 1.8 | 0.4 | 0.08 | 10 |
Body Surface Area (m2) | 0.007 | 0.025 | 0.15 | 0.05 | 0.02 | 0.5 |
Km factor | 3 | 6 | 12 | 8 | 5 | 20 |
Animal A (mg/kg) = Animal B (mg/kg) multiplied by | Animal B Km |
Animal A Km |
For example, to modify the dose of resveratrol used for a mouse (22.4 mg/kg) to a dose based on the BSA for a rat, multiply 22.4 mg/kg by the Km factor for a mouse and then divide by the Km factor for a rat. This calculation results in a rat equivalent dose for resveratrol of 11.2 mg/kg.
Molecular Weight | 487.29 |
Formula | C24H16Cl2O7 |
CAS Number | 4091-99-0 |
Purity | 99.61% |
Solubility | DMSO: ≥ 90 mg/mL |
Storage | -20°C, protect from light |
Redox environment in stem and differentiated cells: A quantitative approach.
Lyublinskaya OG, et al. Redox Biol. 2017 Aug;12:758-769. PMID: 28426982.
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