ROS reactive oxygen species assay kit, as an important biochemical detection tool, is widely used in various fields such as biology, medicine, environmental science, etc. Mainly used to detect the levels of reactive oxygen species (ROS) in cells, tissues, or biological fluids, helping researchers gain a deeper understanding of the effects of oxidative stress on cells and tissues, and thus revealing the association between ROS and disease occurrence and development.
1、 Basic principles
The core principle of ROS reactive oxygen species assay kit is mainly based on fluorescence probe method, among which dichlorodihydrofluorescein acetoacetate (DCFH-DA) or H2DCFDA are commonly used as fluorescence probes. These probes themselves do not possess fluorescent properties, but can freely penetrate the cell membrane and enter the cell. Once inside the cell, it will be hydrolyzed by intracellular esterases, generating DCFH that cannot penetrate the cell membrane. In cells, ROS can specifically oxidize DCFH to generate dichlorofluorescein (DCF) with strong fluorescent properties. By detecting the fluorescence intensity of DCF, the level of intracellular ROS can be indirectly reflected.
2、 Main features
High selectivity: The ROS reactive oxygen species kit can specifically detect ROS, avoiding interference from other chemicals and ensuring the accuracy of measurement results.
High sensitivity: This kit can detect extremely low concentrations of ROS, allowing researchers to accurately measure ROS levels and reveal the impact of small changes on cell function and disease development.
Quick and easy: Provides a fast and convenient method to measure ROS levels, saving laboratory personnel time and effort.
Reliability and repeatability: It has good reliability and repeatability, and can be repeatedly measured under different experimental conditions to ensure the accuracy and repeatability of the results.
3、 Operation steps
When using ROS reactive oxygen species assay kits, the following steps are usually required:
Cell preparation: Ensure that the cell confluence reaches an appropriate range during testing, such as 50%~70%.
Drug induction (optional): Add appropriate concentrations of drugs to induce ROS production according to experimental requirements.
Probe loading: Dilute DCFH-DA or H2DCFDA to an appropriate concentration, add it to the cell culture system, and incubate in the dark for a certain period of time.
Cell cleaning: Remove probes that have not entered the cell to avoid interference from background fluorescence.
Fluorescence detection: Use fluorescence microscopy, flow cytometry and other equipment to detect the fluorescence intensity of DCF and analyze the intracellular ROS level.
4、 Application Fields
ROS reactive oxygen species assay kits have a wide range of applications in biomedical research, including oxidative stress studies, drug screening, environmental toxicity assessment, and more. By measuring ROS levels, researchers can assess the degree of cellular oxidative stress, reveal the association between ROS and disease occurrence and development, and provide scientific basis for disease diagnosis, prevention, and treatment. Meanwhile, the kit can also be used to evaluate the effects of candidate drugs on ROS production and clearance, in order to identify potential antioxidants or oxidants. In the field of environmental science, ROS reactive oxygen species assay kits can be used to evaluate the impact of environmental pollutants on cellular ROS production, revealing the relationship between environmental toxicity and oxidative stress.
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