Overview

  • Product name
    Thioredoxin Reductase 1 (TXNRD1) Activity Assay Kit
    See all Thioredoxin reductase kits
  • Detection method
    Colorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Lysates 4 2.9%
    Inter-assay
    Sample n Mean SD CV%
    Lysates 4 3.2%
  • Sample type
    Cell culture extracts, Tissue Extracts
  • Assay type
    Cell-based (quantitative)
  • Range
    5 µg/ml - 1000 µg/ml
  • Assay duration
    Multiple steps standard assay
  • Species reactivity
    Reacts with: Human
    Does not react with: Mouse, Rat
  • Product overview

    Abcam’s Thioredoxin Reductase 1 (TXNRD1) Activity Assay kit is designed for the sensitive and accurate measurement of Thioredoxin Reductase 1 activity in Human cell and tissue extracts.

    The assay uses a 96-well plate with an antibody specific to isoform 1 of thioredoxin reductase to isolate the isozyme pre-coated onto the wells. Samples are added to the wells and incubated at room temperature, any Thioredoxin Reductase 1 present in the sample will be immobilized in the well. After washing, the Reaction Buffer is added to each well and enzyme activity is measured. By analyzing the enzyme’s activity in an isolated context, outside of the cell and free from other isoforms, an accurate measurement of the enzyme’s functional state can be understood.

  • Notes

    Thioredoxin Reductase 1 (TXNRD1) is a selenium-containing enzyme part of the thioredoxin system responsible for regulating oxidative stress and redox signaling via reduction of disulfide bonds. The dimer, Thioredoxin Reductase 1, reduces thioredoxin and other substrates using NADPH and an FAD cofactor. Thioredoxin Reductase 1 involvement in protecting against oxidative stress and injury, regulation of cellular development and growth, and various other cellular processes make it an interesting target for studies of various cancers, AIDS, and other diseases. Humans express three different isozymes of thioredoxin reductase, isoform 1 is the cytosolic form, isoform 2 is the mitochondrial form, and isoform 3 is a testes specific form.

     

    The enzyme activity is determined by following the NADPH-assisted reduction of 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) to 5-thio-2-nitrobenzoic acid (TNB) which leads to increased absorbance at 412 nm. Two moles of TNB are formed from the oxidation of one mole of NADPH according to the reaction shown below.

     

    DTNB + NADPH + H+ ↔ 2 TNB + NADP+

     

    The molar extinction coefficient of TNB is 14.15 X 103 mM-1cm-1. Thioredoxin Reductase 1 activity is controlled by enzyme amount. An antibody specific to isoform 1 of thioredoxin reductase is used to isolate the enzyme from a bulk protein source for isoform-specific enzymatic activity measurements. Because the activity measured is specific to the isolated enzyme, a thioredoxin reductase inhibitor is not required for background signal subtraction.

  • Platform
    Microplate

Properties

  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Blocking Buffer 1 x 8ml
    10X Buffer 1 x 25ml
    200X DTNB 1 x 110µl
    200X NADPH 1 vial
    5X Activity Solution 1 x 5ml
    Extraction Buffer 1 x 15ml
    TXNRD1 96-well microplate 1 unit
  • Research areas
  • Relevance
    Thioredoxin reductase (TrxR) (EC 1.8.1.9) is a ubiquitous enzyme which is involved in many cellular processes such as cell growth, p53 activity, and protection against oxidation stress, etc. The mammalian TrxR reduces thioredoxins as well as non-disulfide substrates such as selenite, lipoic acids, lipid hydroperoxides, and hydrogen peroxide.
  • Cellular localization
    TXNRD1: Cytoplasmic. TXNRD2: Cytoplasmic. Nuclear. Microsome. Endoplasmic reticulum. TXNRD3: Mitochondrial.
  • Alternative names
    • EC 1.8.1.9
    • gene associated with retinoic and interferon-induced mortality 12 protein
    • Gene associated with retinoid IFN induced mortality 12 protein
    • GRIM 12
    • KM 102 derived reductase like factor
    • KM-102-derived reductase-like factor
    • MGC9145
    • oxidoreductase
    • Selenoprotein Z
    • SELZ
    • TGR
    • Thioredoxin and glutathione reductase
    • Thioredoxin glutathione reductase
    • Thioredoxin reductase 1
    • thioredoxin reductase 1, cytoplasmic
    • Thioredoxin reductase 2
    • thioredoxin reductase 2, mitochondrial
    • Thioredoxin reductase 3
    • Thioredoxin reductase beta
    • Thioredoxin reductase GRIM 12
    • thioredoxin reductase TR1
    • thioredoxin reductase TR2
    • thioredoxin reductase TR3
    • TR
    • TR beta
    • TR1
    • TR2
    • TR3
    • TrxR
    • TRXR1
    • TRXR2
    • TRXR3
    • TXNR
    • TXNRD1
    • TXNRD2
    • TXNRD3
    see all
  • Database links

Images

  • The data shown above was collected at the endpoint after 30 minutes.

  • After the rate/slope of each lane is extracted from the linear range of the time point data, it is expressed as rate (mOD/min) per microgram of cell lysate added per well. The extinction for the DTNB dye is 9.9/ mM / well.

Protocols

References

This product has been referenced in:
  • Xiaobo C  et al. TUSC2(FUS1)-erlotinib Induced Vulnerabilities in Epidermal Growth Factor Receptor(EGFR) Wildtype Non-small Cell Lung Cancer(NSCLC) Targeted by the Repurposed Drug Auranofin. Sci Rep 6:35741 (2016). Read more (PubMed: 27845352) »

See 1 Publication for this product

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