There are many methods to study the functions of antioxidants, and according to whether the organism participates in the experiment, the methods can be divided into in vitro and in vivo assays. The in vitro assay is simple in operation and fast in detection, which is widely used in production practice. However, the method has limitations, including contradictory results, and lack of a uniform standard to precisely quantify the extent of antioxidant capacity. Moreover, because of the low biological correlations among these methods, measuring results cannot reflect the biological antioxidants in the body. The biocorrelation of the in vivo assay is higher and the results are more reliable. Therefore, studying the function of antioxidants using C. elegans is an efficient and low-cost in vivo method can be applied to detect reactive oxygen species (ROS) scavenging abilities, the content of antioxidant enzymes and the aspect of DNA oxidative damage.
As a leading service provider in C. elegans model service, Creative Biogene' sophisticated equipment, advanced technologies and highly experienced staff are available to provide our customers with the comprehensive C. elegans antioxidant function assay service, including detection of ROS, oxidative damage products, related enzyme activity. With the help of Creative Biogene, you can improve research outcomes and dramatically improve the speed of success. We are committed to offering customers fast turnaround, high-quality services at competitive prices.
Determination of the ROS Level
During the cell metabolism process, a series of ROS are generated, such as superoxide anion, hydroxyl radicals, and hydrogen peroxide. While affecting lipids, membranes, and DNA, ROS-induced oxidative damage can disrupt proteostasis especially. Additionally, high concentrations of ROS can induce cell apoptosis. Therefore, determination of the ROS Level is essential in antioxidant function assay. Here we choose fluorescence analysis, which is widely and directly used to detect intracellular ROS levels. Fluorescence analysis has many advantages, such as high sensitivity, good selectivity, and an extremely wide dynamic response range. Moreover, it is relatively simple and economical.
Detection of Oxidative Damage Products
The oxidative damage products refer to adducts produced by ROS damaging biological macromolecules such as DNA, proteins and lipids. Among them, urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG), an oxidative DNA damage adduct, is commonly analyzed as an excellent marker of DNA lesions. CE-LIF detection involves the use of a specific antibody to detect the DNA lesion (8-OHdG) and consecutive fluorescence labeling, and the lesion of interest was detected using a fluorescence detector. The method has great advantages, such as high separation efficiency, good selectivity, low limit of detection, simplicity and low cost of analysis.
Measurement of Antioxidant Enzyme Activity
Oxidative stress-induced ROS quickly accumulates in cells, leading to oxidative damage of tissue, and consequently, accelerated death of organisms. In vivo, antioxidant enzymes including Catalase (CAT), Superoxide Dismutase (SOD) play a crucial role in protecting the organ system against over-production of ROS. SOD is an important antioxidant enzyme, and has been documented to play an important role in catalyzing the dismutation of superoxide into water and H2O2. We provide fluorescence analysis for quantitative detection of activity of SOD using fluorescence labeling instruments. CAT is considered as the principal H2O2 scavenging enzyme, catalyzing the H2O2 into H2O and O2. Iodimetric titration and ultraviolet spectrophotometry are combined to measure the activity of CAT.
* For research use only.