PRX Pathway (TRX, TXNRD1, PRX1) Western Blot Cocktail (ab184868)
Key features and details
- Sample type: Cell Lysate
製品の概要
-
製品名
PRX Pathway (TRX, TXNRD1, PRX1) Western Blot Cocktail -
サンプルの種類
Cell Lysate -
種交差性
交差種: Mouse, Rat, Human -
製品の概要
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.
-
特記事項
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 -
アプリケーション
適用あり: WBmore details
製品の特性
-
保存方法
Store at -20°C. Please refer to protocols. -
内容 200 µl 250X PRX Pathway (TRX, TXNRD1, PRX1) Western Blot Cocktail 1 x 200µl -
研究分野
-
参照データベース
- 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
see all
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab184868の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
| アプリケーション | Abreviews | 特記事項 |
|---|---|---|
| WB |
Use at an assay dependent concentration.
-WB samples should be heated to 95°C for 10 minutes in sample buffer containing DTT before loading. -Suggested working concentration is 1X for the antibody cocktail. -Suggested incubation buffer is 5% milk in TBS + 0.05%Tween 20.
Optimized for cell lysates, may also work on tissue homogenates.
Expected and observed MWs: Thioredoxin: 12 kDa Thioredoxin Reductase 1: 55 kDa Peroxiredoxin 1: 22 kDa |
| 特記事項 |
|---|
|
WB
Use at an assay dependent concentration. -WB samples should be heated to 95°C for 10 minutes in sample buffer containing DTT before loading. -Suggested working concentration is 1X for the antibody cocktail. -Suggested incubation buffer is 5% milk in TBS + 0.05%Tween 20.
Optimized for cell lysates, may also work on tissue homogenates.
Expected and observed MWs: Thioredoxin: 12 kDa Thioredoxin Reductase 1: 55 kDa Peroxiredoxin 1: 22 kDa |
画像
-
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
-
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
-
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.
プロトコール
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.
データシートおよび資料
-
SDS download
-
Datasheet download
参考文献 (5)
ab184868 は 5 報の論文で使用されています。
- von Mässenhausen A et al. Dexamethasone sensitizes to ferroptosis by glucocorticoid receptor-induced dipeptidase-1 expression and glutathione depletion. Sci Adv 8:eabl8920 (2022). PubMed: 35108055
- Trstenjak Prebanda M et al. Altered Expression of Peroxiredoxins in Mouse Model of Progressive Myoclonus Epilepsy upon LPS-Induced Neuroinflammation. Antioxidants (Basel) 10:N/A (2021). PubMed: 33673502
- Trstenjak Prebanda M et al. Upregulation of Mitochondrial Redox Sensitive Proteins in LPS-Treated Stefin B-Deficient Macrophages. Cells 8:N/A (2019). PubMed: 31766320
- Trewin AJ et al. Acute exercise alters skeletal muscle mitochondrial respiration and H2O2 emission in response to hyperinsulinemic-euglycemic clamp in middle-aged obese men. PLoS One 12:e0188421 (2017). PubMed: 29161316
- Bhatt NM et al. Restoring redox balance enhances contractility in heart trabeculae from type 2 diabetic rats exposed to high glucose. Am J Physiol Heart Circ Physiol 308:H291-302 (2015). WB ; Rat . PubMed: 25485897