Product nameThioredoxin Reductase 1 (TXNRD1) Activity Assay Kit
See all Thioredoxin reductase kits
Intra-assay Sample n Mean SD CV% Lysates 4 2.9% Inter-assay Sample n Mean SD CV% Lysates 4 3.2%
Sample typeCell culture extracts, Tissue Extracts
Assay typeCell-based (quantitative)
Range5 µg/ml - 1000 µg/ml
Species reactivityReacts with: Human
Does not react with: Mouse, Rat
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.
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.
Storage instructionsStore 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
RelevanceThioredoxin reductase (TrxR) (EC 126.96.36.199) 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 localizationTXNRD1: Cytoplasmic. TXNRD2: Cytoplasmic. Nuclear. Microsome. Endoplasmic reticulum. TXNRD3: Mitochondrial.
- EC 188.8.131.52
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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) »