Recombinant Human Glyoxalase II Protein, CF

R&D Systems | Catalog # 5944-GO

R&D Systems
100% Guarantee

Key Product Details

  • R&D Systems E. coli-derived Recombinant Human Glyoxalase II Protein (5944-GO)
  • 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

NP_001035517

Applications

Enzyme Activity
View all Glyoxalase II Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human Glyoxalase II protein
Met1-Asp260, with a C-terminal 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

Met1

Predicted Molecular Mass

30 kDa

SDS-PAGE

29-30 kDa, reducing conditions

Activity

Measured by its ability to hydrolyze S-lactoylglutathione.
The specific activity is >25,000 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

5944-GO
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol.
Shipping The product is shipped with dry ice or equivalent. 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, -70 °C as supplied.
  • 3 months, -70 °C under sterile conditions after opening.

Background: Glyoxalase II

The glyoxalase system consists of two enzymes, Glyoxalase I and Glyoxalase II. The purpose of the glyoxalase system is to detoxify alpha -ketoaldehydes, particularly methylglyoxal, a by-product of glycolysis (1). Glyoxalase II catalyzes the hydrolysis of S-lactoylglutathione, the product of the glyoxalase I reaction, to glutathione and lactate (2). Glyoxalase II exists as two isoforms, one cytosolic and one mitochondrial (3). The enzyme is believed to be important for the response of cells to stress (4).

References

  1. Thornalley, P.J. (1996) Gen. Pharmacol. 27:565.
  2. Ridderström, M. et al. (1996) J. Biol. Chem. 271:319.
  3. Cordell, P.A. et al. (2004) J. Biol. Chem. 279:28653.
  4. Xu, Y. and X. Chen (2006) J. Biol. Chem. 281:26702.

Alternate Names

GLX2, HAGH

Entrez Gene IDs

3029 (Human); 14651 (Mouse); 24439 (Rat)

Gene Symbol

HAGH

UniProt

NP_001035517

Additional Glyoxalase II Products

Product Documents for Recombinant Human Glyoxalase II Protein, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Recombinant Human Glyoxalase II Protein, CF

Coomassie is a registered trademark of Imperial Chemical Industries Ltd.

For research use only

Related Research Areas

Redox Enzymes

Citations for Recombinant Human Glyoxalase II Protein, CF

Customer Reviews for Recombinant Human Glyoxalase II Protein, CF

There are currently no reviews for this product. Be the first to review Recombinant Human Glyoxalase II Protein, CF and earn rewards!

Have you used Recombinant Human Glyoxalase II Protein, CF?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review
Amazon Gift Card

Protocols

View specific protocols for Recombinant Human Glyoxalase II Protein, CF (5944-GO):

Materials
  • Assay Buffer: 50 mM Tris, 250 mM NaCl, pH 7.5
  • Recombinant Human Glyoxalase II (rhGlyoxalase II) (Catalog # 5944-GO)
  • Substrate: S-Lactoylglutathione (Sigma, Catalog # L7140), 100 mM stock in deionized water
  • 5, 5’-dithiobis(2-nitrobenzoic acid) (DTNB) (Sigma, Catalog # D8130), 10 mM stock in DMSO
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhGlyoxalase II to 0.4 ng/µL in Assay Buffer.
  2. Dilute Substrate to 2 mM in Assay Buffer with 400 µM DTNB to form Substrate Mixture.
  3. Load 50 µL of 0.4 ng/µL rhGlyoxalase II into a plate and start the reaction by loading 50 µL of Substrate Mixture. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of Substrate Mixture.
  4. Read at 405 nm (absorbance) in kinetic mode for 5 minutes.
  5. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (OD/min) 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 13260 M-1cm-1 
     ***Using the path correction 0.320 cm
     Note: the output of many spectrophotometers is in mOD. Per Well:
  • rhGlyoxalase II: 0.020 µg
  • S-Lactoylglutathione: 1 mM
  • DTNB: 200 µM

FAQs

No product specific FAQs exist for this product.

View all FAQs for Proteins and Enzymes