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Principle and Operating Steps of ROS Reactive Oxygen Species Kit
Reactive oxygen species (ROS), as an important class of active molecules in organisms, play critical roles in cell signaling, immune response, and pathological processes. In order to accurately detect the level of intracellular ROS, a ROS reactive oxygen species assay kit has been developed, which has wide application value in multiple fields such as biology, medicine, and environmental science. This article introduces the basic principles, application areas, and operating steps of ROS reactive oxygen species assay kits.
1、 Basic principles
The core principle of ROS reactive oxygen species assay kit is mainly based on fluorescence probe method, among which the commonly used method is to use dichlorodihydrofluorescein acetoacetate (DCFH-DA) as a fluorescence probe. DCFH-DA itself does not possess fluorescent properties, but it can freely penetrate the cell membrane and enter the cell. In cells, DCFH-DA is hydrolyzed by esterases to produce DCFH, which is effectively retained as a "sensor" for ROS detection because it cannot cross the cell membrane again. When intracellular ROS (such as superoxide anions, hydrogen peroxide, and hydroxyl radicals) react with DCFH, dichlorofluorescein (DCF) with strong fluorescent properties is generated. By detecting the fluorescence intensity of DCF, the level of intracellular ROS can be indirectly reflected.
2、 Application Fields
ROS reactive oxygen species assay kits have broad application prospects in multiple fields:
Cell biology research: used to monitor the changes in ROS levels of different cell types under physiological or pathological conditions, and to reveal the mechanisms of ROS in cell proliferation, differentiation, apoptosis, and other processes.
Oxidative stress and disease research: Evaluate the role of oxidative stress in the occurrence and development of cardiovascular diseases, neurodegenerative diseases, cancer, etc., and provide scientific basis for disease diagnosis, prevention, and treatment.
Drug screening and evaluation: In the process of drug development, the impact of drugs on intracellular ROS levels is detected to evaluate the antioxidant capacity or potential toxicity of drugs, providing important references for drug screening and optimization.
Environmental and Health Research: Study the impact of environmental factors (such as pollutants, radiation, etc.) on human cellular ROS levels, and evaluate the potential harm of environmental pollution to health.
3、 Operation steps
When using ROS reactive oxygen species assay kits, the following steps are usually followed:
Cell preparation: Ensure that the cell confluence reaches the appropriate range during testing.
Probe loading: Dilute DCFH-DA 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.
In summary, ROS reactive oxygen species assay kit is an efficient and accurate tool for detecting intracellular ROS levels, and has important value in scientific research and practical applications.