Description
Key facts
Applications:WBSee reactivity dataSee the reactivity data table below for information on validated species and application combinations.,
Target:CatalaseSee target data,
Reacts with:Human,
Form:LiquidSee storage information,
Storage buffer:Preservative: 0.01% Sodium azideConstituents: 50% Glycerol (glycerin, glycerine), 0.9% Sodium chloride, 0.6% Tris, 0.05% BSA
Product details:
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 Down's 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 95°C 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.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-+4°C, Appropriate long-term storage conditions--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
The oxidative stress defense proteins protect cells from damage induced by reactive oxygen species (ROS). These proteins including superoxide dismutase catalase and glutathione peroxidase counteract oxidative stress by neutralizing harmful ROS. Superoxide dismutase with a mass of approximately 32 kDa is expressed in the cytoplasm mitochondria and extracellular space across various tissues. Catalase and glutathione peroxidase play similar roles ensuring a balance is maintained in cellular environments prone to oxidative reactions.
Biological function summary
These proteins engage with multiple cell processes to maintain cellular health by preventing oxidative damage to DNA proteins and lipids. These defense proteins often form part of larger complexes that work synergistically to reduce oxidative stress levels. By participating in antioxidant defense they limit cellular malfunction and support normal cellular signaling and function encompassing broad cellular protective mechanisms.
Pathways
Proteins involved in oxidative stress defense play a role in the detoxification pathway reducing oxidative stress markers and maintaining cellular redox balance. The pathway involves interactions with key endogenous antioxidants like glutathione. They also intersect with inflammation-related pathways wherein oxidative stress-related damage often triggers inflammatory responses. Oxidative stress defense proteins may also interact with the nuclear factor erythroid 2-related factor 2 (Nrf2) a transcription factor important for inducing antioxidant response elements.
Oxidative stress defense proteins have connections to conditions such as neurodegenerative diseases like Alzheimer's and cardiovascular disorders. In Alzheimer's disease abnormal oxidation affects amyloid-beta proteins where oxidative stress exacerbates symptoms. Additionally oxidative stress influences pathways contributing to plaque formation in cardiovascular disease suggesting a role in disease progression. These proteins’ functions strongly tie to proteins like amyloid precursor protein in Alzheimer’s and low-density lipoprotein in cardiovascular disease emphasizing their critical role in disease pathophysiology.
