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Cat. No.: SAK-100 (for 100T)

Description

Superoxide Assay Kit is a reagent kit used for the rapid and highly sensitive detection of superoxide radicals. This assay kit utilizes the ability of superoxide radicals to reduce WST-1, resulting in the production of a soluble colored compound. The detection reagent in this assay kit contains Catalase, which can eliminate interference from hydrogen peroxide and other peroxides on the color development of WST-1, ensuring more accurate results. Additionally, the assay kit provides Superoxide Dismutase (SOD) to validate the presence of superoxide radicals detected by the kit, thereby excluding potential interference from other reagents used in the assay system. The addition of SOD to this assay kit typically inhibits more than 90% of superoxide radicals.

WST-1 is a compound similar to MTT and can be reduced by superoxide radicals to form an orange-colored formazan product. The more superoxide radicals are generated, the faster the color develops, leading to higher absorbance values obtained in the measurement.

Using WST-1 as the detection reagent offers several advantages over traditional cytochrome c-based methods for detecting superoxide radicals. Firstly, WST-1 itself has very low absorbance (typically around OD450, depending on the instrument and microplate), while cytochrome c has high absorbance (typically around OD550). Therefore, compared to cytochrome c, WST-1 provides much higher detection sensitivity. Secondly, the reduction product of WST-1 is stable and soluble, with minimal toxicity to cells, making the WST-1 method more suitable for high-throughput screening. Moreover, due to the limitations in sensitivity, the cytochrome c method is generally not suitable for 96-well plate measurements, whereas the WST-1 method can be used with a 96-well plate, enabling more convenient detection. A comparison of the effects of using the WST-1 or cytochrome c method for superoxide radical detection can be seen in Figure 1. In Figure 1, neutrophils at a concentration of 500,000 cells/mL were stimulated with PMA at 37°C for a specified time, and measurements were taken using the WST-1 or cytochrome c method. The WST-1 method measures OD450, while the cytochrome c method measures OD550. Furthermore, compared to the luminol method, the WST-1 method exhibits similar sensitivity but only requires a regular microplate reader without the need for a luminometer, resulting in faster detection.

Superoxide radicals typically refer to the superoxide anion (O2.-), which is a reactive oxygen species. In the respiratory chain, NADPH oxidase transfers electrons to oxygen molecules, leading to the production of superoxide anions (O2.-). Superoxide anions (O2.-) are powerful oxidants and can be generated by stimulated white blood cells to combat microbial infections and other threats. However, superoxide anions (O2.-) can also cause oxidative damage and are closely associated with the development of various diseases.

This assay kit can test 100 samples.

Additional Notes