Oxidative Stress Defense (Catalase, SOD1, TRX, smooth muscle Actin) Western Blot Cocktail

Description

This Oxidative Stress Defense Western Blot Cocktail is designed to determine the relative abundance of several important proteins involved in the protection of cells against oxidative stress and the regulation of reactive oxygen species ROS . Reactive oxygen species are produced naturally in cells as byproducts of the metabolism of oxygen as well as in response to various environmental stresses including UV radiation, pollutants, and heat exposure. Additionally, ROS levels can be altered by disease and injury, including cancer, neurodegenerative disease, cardiovascular disease, ischemia, stroke and aging. Reactive oxygen species also play an important role in cell signaling, a process called redox signaling. The regulation of ROS within cells is important for maintaining a proper homeostasis. Superoxide dismutase 1 SOD1 scavenges harmful superoxides O2- within cells protecting them from harmful oxidation of lipids, proteins and nucleic acids. Its altered expression levels have been linked to Downs syndrome, ALS and various cancers. Similarly, the hydrogen peroxide H2O2 scavenging enzyme, catalase, also regulated ROS concentrations within cells by reducing H202 into less reactive O2 and water. Thioredoxin is a small enzyme 12kDa that facilitates the reduction of other enzymes via cysteine thiol-disulfide exchange. Thioredoxin is used by cells to reduce ROS amounts and in redox signaling processes. Finally, alpha smooth muscle actin was included in the cocktail as a loading control. Widely expressed, smooth muscle actin is involved in cell structure and motility. These four readouts are easily resolved by western blot given their different molecular weights. Because they are all rabbit monoclonal antibodies, an anti-rabbit secondary should be used for detection. Expected and observed MWs: Catalase: 60 kDa Smooth Muscle Actin: 42 kDa Superoxide Dismutase 1: 16 kDa Thioredoxin: 12 kDa WB Notes: The provided 250X antibody cocktail should be diluted to 1X in appropriate dilution buffer before use. Suggested dilution buffer is 5 % milk/PBS+0.05 %Tween 20. WB samples should be heated to 95C for 5 minutes in sample buffer before loading. The cocktail contains 50 % glycerol, can be stored at -20C. No aliquoting necessary. Related products Review the oxidative stress marker and assay guide to learn about more assays for oxidative stress.