PRX Pathway (TRX, TXNRD1, PRX1) Western Blot Cocktail (ab184868)
Key features and details
- Sample type: Cell Lysate
Overview
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Product name
PRX Pathway (TRX, TXNRD1, PRX1) Western Blot Cocktail -
Sample type
Cell Lysate -
Species reactivity
Reacts with: Mouse, Rat, Human -
Product overview
The PRX pathway Western blot cocktail is designed to determine the relative abundance of several important proteins involved in the thioredoxin redox pathway.
Thioredoxin (TRX) 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. Thioredoxin reductase 1 (TXNRD1) is found in the cytoplasm and possesses glutaredoxin and thioredoxin reductase activity. TXNRD1 utilizes its redox-active disulfide bond to reduce oxidized thioredoxin. Also involved in the thioredoxin redox pathway and additionally shown to be upregulated in a variety of cancers is the relatively abundant peroxiredoxin 1 (PRX1). Peroxiredoxin 1 is an antioxidant enzyme that helps regulate peroxide levels and is involved in redox signaling. Peroxiredoxin 1’s activity is regenerated after being reduced by thioredoxin.
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Notes
These three readouts are easily resolved by Western blot given their different molecular weights. As all primary antibodies in the WB cocktail are rabbit antibodies, an anti-rabbit secondary should be used for detection.
Expected and observed MWs:
Thioredoxin: 12 kDa
Thioredoxin Reductase 1: 55 kDa
Peroxiredoxin 1: 22 kDa -
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 PRX Pathway (TRX, TXNRD1, PRX1) Western Blot Cocktail 1 x 200µl -
Research areas
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Database links
- Entrez Gene: 5052 Human
- Entrez Gene: 7295 Human
- Entrez Gene: 7296 Human
- Entrez Gene: 18477 Mouse
- Entrez Gene: 50493 Mouse
- Entrez Gene: 117254 Rat
- Entrez Gene: 58819 Rat
- SwissProt: P10599 Human
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Images
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All three protein targets resolved separately and in the combined cocktail on HeLa cell lysate.
WB lysate sample was heated at 95°C for 10 minutes before loading. Performed under reducing conditions. All blocking and antibody incubation steps were done in 5% milk in TBST. Developed using the ECL technique.
Exposure time: 1 minute.
Sample: HeLa Cell Lysate – 20 µg/lane
Lane 1: Anti-Thioredoxin Reductase 1 antibody
Lane 2: Anti-Peroxiredoxin 1 antibody
Lane 3: Anti-Thioredoxin antibody
Lane 4: PRX Pathway WB Cocktail
Secondary: HRP-conjugated Anti-Rabbit IgG
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Western blot analysis of multiple cell lines using the PRX Pathway Western Blot Cocktail.The PRX Pathway was used on various cell types to determine the relative amounts of thioredoxin reductase 1, peroxiredoxin 1, and thioredoxin.
All blocking and antibody incubation steps were done in 5% milk in TBST. Developed using the ECL technique.
Exposure time: 1 minute.
20 µg of each cell lysate was loaded per lane after heating for 10 minutes at 95°C.
Lane 1: Hela
Lane 2: HepG2
Lane 3: HDFn
Lane 4: Hek293T
Lane 5: MCF7
Lane 6: Jurkat
Lane 7: HL60
Lane 8: H9C2
Lane 9: MEF
Secondary: HRP-conjugated Anti-Rabbit IgG
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Relative abundances of target proteins.Densiometric analysis of a Western blot obtained with the PRX Pathway WB cocktail using various cell lines to determine the relative amounts of thioredoxin reductase 1, peroxiredoxin 1, and thioredoxin.
