Recombinant Human Glutathione Reductase Protein, CF

R&D Systems | Catalog # 8866-GR

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Key Product Details

  • R&D Systems E. coli-derived Recombinant Human Glutathione Reductase Protein (8866-GR)
  • Quality control testing to verify active proteins with lot specific assays by in-house scientists
  • All R&D Systems proteins are covered with a 100% guarantee

Source

E. coli

Accession Number

P00390

Applications

Enzyme Activity
View all Glutathione Reductase Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human Glutathione Reductase protein
Met44-Arg522, with an N-terminal Met and 6-His tag

Purity

>85%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Met

Predicted Molecular Mass

53 kDa

SDS-PAGE

53 kDa, reducing conditions

Activity

Measured by its ability to reduce oxidized glutathione to glutathione in an NADPH-dependent manner.
The specific activity is >15,000 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

8866-GR
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, DTT and Glycerol.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: Glutathione Reductase

Glutathione Reductase (GR) is a homodimeric flavoprotein disulfide oxidoreductase that plays a critical role in the prevention of oxidative damage within the cell (1, 2). Glutathione Reductase helps to maintain appropriate levels of intracellular reduced glutathione (GSH) which is essential for maintaining the normal structure of red blood cells and for keeping hemoglobin in the ferrous state (3). Glutathione Reductase together with its co-factor, NADPH, catalyzes the reduction of oxidized glutathione (glutathione disulfide, GSSG) to glutathione. GSH is also a reactant for Glutathione Peroxidase, which converts hydrogen peroxide hydrogen peroxide (H2O2) into water (4). Glutathione Reductase is expressed as both mitochondrial and cytosolic isoforms, as determined by the presence or absence of an N-terminal transit peptide. Human Glutathione Reductase shares 86% and 76% amino acid sequence identity with mouse and rat Glutathione Reductase, respectively (5, 6). Alternative splicing of human Glutathione Reductase generates additional isoforms with deletions in the central pyridine nucleotide-disulfide oxidoreductase domain (7).

References

  1. Deponte, M. (2013) Biochim. Biophys. Acta 1830:3217.
  2. Fujii, J. et al. (2011) J. Clin. Biochem. Nutr. 49:70.
  3. Tsantes, A.E. et al. (2006) Antioxid. Redox Signal. 8:1205.
  4. Lubos, E. et al. (2011) Antioxid. Redox Signal. 15:1957.
  5. Tutic, M. et al. (1990) Eur. J. Biochem. 188:523.
  6. Krauth-Siegel, R.L. et al. (1982) Eur. J. Biochem. 121:259.
  7. Satoh, N. et al. (2010) Biochem. Genet. 48:816.

Alternate Names

GLUR, GRase, GRD1, GSR, HEL-752

Entrez Gene IDs

2936 (Human); 14782 (Mouse); 116686 (Rat)

Gene Symbol

GSR

UniProt

P00390

Additional Glutathione Reductase Products

Product Documents for Recombinant Human Glutathione Reductase Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human Glutathione Reductase Protein, CF

For research use only

Related Research Areas

Redox Enzymes

Citations for Recombinant Human Glutathione Reductase Protein, CF

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Protocols

View specific protocols for Recombinant Human Glutathione Reductase Protein, CF (8866-GR):

Materials
  • Assay Buffer: 0.1 M Tris, pH 7.5
  • Recombinant Human Glutathione Reductase (rhGSR) (Catalog # 8866-GR)
  • NADPH (Sigma, Catalog # N7505), 10 mM stock in deionized water
  • Oxidized Glutathione (Amresco, Catalog # 0524), 50 mM stock in deionized water
  • 96-well Clear Plate (Catalog # DY990)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Create Substrate Mixture containing 0.2 mM NADPH and 1 mM Oxidized Glutathione in Assay Buffer.
  2. Dilute rhGSR to 0.4 µg/mL in Assay Buffer.
  3. Load 50 µL of the 0.4 µg/mL rhGSR to the plate, and start the reaction by adding 50 µL of Substrate Mixture. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of Substrate Mixture.
  4. Read plate in kinetic mode for 5 minutes at an absorbance of 340 nm.
  5. The specific activity can be calculated using the following equation:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (OD/min) x -1 x well volume (L) x 1012 pmol/mol
ext. coeff** (M-1cm-1) x path corr.*** (cm) x amount of enzyme (µg)

      
      *Adjusted for Substrate blank
      **Using the extinction coefficient 6220 M-1cm-1
      ***Using the path correction 0.32 cm
      Note: the output of many spectrophotometers is in mOD.

Per Well:

  • rhGSR: 0.02 µg
  • NADPH: 0.1 mM
  • Oxidized Glutathione: 0.5 mM

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