Recombinant Human Glyoxalase II Protein, CF

Catalog # Availability Size / Price Qty
5944-GO-010
R&D Systems Recombinant Proteins and Enzymes
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Citations (1)
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Recombinant Human Glyoxalase II Protein, CF Summary

Product Specifications

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.
Activity
Measured by its ability to hydrolyze S-lactoylglutathione. The specific activity is >25,000 pmol/min/μg, as measured under the described conditions.
Source
E. coli-derived human Glyoxalase II protein
Met1-Asp260, with a C-terminal 6-His tag
Accession #
N-terminal Sequence
Analysis
Met1
Predicted Molecular Mass
30 kDa
SDS-PAGE
29-30 kDa, reducing conditions

Product Datasheets

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

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.

Assay Procedure

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
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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.
Entrez Gene IDs
3029 (Human); 14651 (Mouse); 24439 (Rat)
Alternate Names
EC 3.1.2.6; GLO2hydroxyacyl glutathione hydrolase; Glx II; GLX2; GLXII; Glyoxalase II; HAGH; HAGH1GLX2; hydroxyacylglutathione hydrolase; hydroxyacylglutathione hydrolase, mitochondrial; hydroxyacylglutathione hydroxylase

Product Specific Notices

Coomassie is a registered trademark of Imperial Chemical Industries Ltd.

Citation for Recombinant Human Glyoxalase II Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

  1. Discovery of a Novel Metallo-�-Lactamase Inhibitor, which can Potentiate Meropenem Activity against Carbapenem-Resistant Enterobacteriaceae
    Authors: M Everett, N Sprynski, A Coelho, J Castandet, M Bayet, J Bougnon, C Lozano, DT Davies, S Leiris, M Zalacain, I Morrissey, S Magnet, K Holden, P Warn, F De Luca, JD Docquier, M Lemonnier
    Antimicrob. Agents Chemother., 2018;0(0):.
    Species: Human
    Sample Types: Chemical Compound
    Applications: Bioassay

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