PRX Pathway (TRX, TXNRD1, PRX1) Western Blot Cocktail (ab184868)

Overview

  • 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.

  • 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

Applications

Our Abpromise guarantee covers the use of ab184868 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.

-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

Images

  • 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

  • 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

  • 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.

References

This product has been referenced in:
  • 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 . Read more (PubMed: 25485897) »

See 1 Publication for this product

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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