Oxidative Stress Defense (Catalase, SOD1, TRX, smooth muscle Actin) Western Blot Cocktail (ab179843)
Key features and details
- Sample type: Cell Lysate, Tissue Homogenate
Overview
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Product name
Oxidative Stress Defense (Catalase, SOD1, TRX, smooth muscle Actin) Western Blot Cocktail -
Sample type
Cell Lysate, Tissue Homogenate -
Species reactivity
Reacts with: Human
Does not react with: Mouse, Rat -
Product overview
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:
- - WB samples should be heated to 95°C for 5 minutes in sample buffer before loading.
- - Suggested working concentration is 1X for primary antibody cocktail.
- - Suggested dilution buffer is 5% milk/PBS+0.05%Tween 20.
The cocktail contains 50% glycerol, can be stored at -20C. No aliquoting necessary.
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Notes
Related products
Review the oxidative stress marker and assay guide to learn about more assays for oxidative stress.
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Tested applications
Suitable for: WBmore details
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 200 µl 250X Oxidative Stress Defense WB cocktail 1 x 200µl -
Research areas
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Alternative names
- AAT6
- ACTA2
- actin, alpha 2, smooth muscle, aorta
see all -
Database links
- Entrez Gene: 847 Human
- Entrez Gene: 59 Human
- Entrez Gene: 6647 Human
- Entrez Gene: 7295 Human
- Omim: 11500 Human
- Omim: 102620 Human
- Omim: 147450 Human
- Omim: 187700 Human
see all
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab179843 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB |
Use at an assay dependent concentration.
Antibody cocktail should be diluted to 1X in appropriate dilution buffer before use. WB samples should be heated to 95°C for 5 minutes in sample buffer before loading. Suggested dilution buffer is 5% milk/PBS+0.05%Tween 20. |
| Notes |
|---|
|
WB
Use at an assay dependent concentration. Antibody cocktail should be diluted to 1X in appropriate dilution buffer before use. WB samples should be heated to 95°C for 5 minutes in sample buffer before loading. Suggested dilution buffer is 5% milk/PBS+0.05%Tween 20. |
Images
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Relative abundances of target proteins normalized to Smooth Muscle Actin control signal.
Densiometric analysis of a western blot using ab179843 was used on various cell types to determine the relative amounts of catalase, superoxide dismutase 1 and thioredoxin.
25 ug of each cell lysate was loaded per lane after heating for 5 minutes at 95°C.
Lane 1: HepG2
Lane 2: HeLa
Lane 3: HDFn
Lane 4: HL60
Lane 5: Jurkat
Lane 6: MCF7
Lane 7: Hek293T
Secondary: HRP-conjugated Anti-Rabbit IgG
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All four protein targets resolved separately and in the combined cocktail on HepG2 cell lysate.WB lysate sample was heated at 95°C for 5 minutes before loading. Performed under reducing conditions.
All blocking and antibody incubation steps were done in 5% milk in PBST.
Developed using the ECL technique.
Exposure time: 1 minute.
Sample: HepG2 Cell Lysate – 25 µg/lane
Lane 1: Anti-Catalase antibody
Lane 2: Anti-Smooth muscle actin antibody
Lane 3: Anti-Superoxide dismutase 1 antibody
Lane 4: Anti-Thioredoxin antibody
Lane 5: ab179843 Oxidative Stress Defense WB Cocktail
Secondary: HRP-conjugated Anti-Rabbit IgG
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (8)
ab179843 has been referenced in 8 publications.
- Huntosova V et al. Influence of Oxidative Stress on Time-Resolved Oxygen Detection by [Ru(Phen)3]2+ In Vivo and In Vitro. Molecules 26:N/A (2021). PubMed: 33477558
- Nylund P et al. A distinct metabolic response characterizes sensitivity to EZH2 inhibition in multiple myeloma. Cell Death Dis 12:167 (2021). PubMed: 33579905
- Nowaczyk M et al. Addition of Popular Exogenous Antioxidant Agent, PBN, to Culture Media May Be an Important Step to Optimization of Myogenic Stem/Progenitor Cell Preparation Protocol. Antioxidants (Basel) 10:N/A (2021). PubMed: 34203726
- Bishop A et al. Fluorescent tools to analyse peroxisome-ER interactions in mammalian cells. Contact (Thousand Oaks) 2:N/A (2019). PubMed: 31198905
- Keatley K et al. Integrated Approach Reveals Role of Mitochondrial Germ-Line Mutation F18L in Respiratory Chain, Oxidative Alterations, Drug Sensitivity, and Patient Prognosis in Glioblastoma. Int J Mol Sci 20:N/A (2019). PubMed: 31323957
- Lo Dico A et al. Intracellular Redox-Balance Involvement in Temozolomide Resistance-Related Molecular Mechanisms in Glioblastoma. Cells 8:N/A (2019). PubMed: 31653091
- Rendleman J et al. New insights into the cellular temporal response to proteostatic stress. Elife 7:N/A (2018). PubMed: 30272558
- Svarcbahs R et al. Removal of prolyl oligopeptidase reduces alpha-synuclein toxicity in cells and in vivo. Sci Rep 8:1552 (2018). PubMed: 29367610