Recombinant Human 6-Phosphogluconate Dehydrogenase/PGD, CF

R&D Systems | Catalog # 8964-DH

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Key Product Details

  • R&D Systems E. coli-derived Recombinant Human 6-Phosphogluconate Dehydrogenase/PGD (8964-DH)
  • 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

P52209

Applications

Enzyme Activity
View all 6-Phosphogluconate Dehydrogenase/PGD Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human 6-Phosphogluconate Dehydrogenase/PGD protein
Ala2-Ala483, with a N-terminal Met and 6-His tag

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

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

N-terminal Sequence Analysis

Met

Predicted Molecular Mass

54 kDa

SDS-PAGE

48 kDa, reducing conditions

Activity

Measured by its ability to dehydrogenate 6-phosphogluconic acid.
The specific activity is >6000 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

8964-DH
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, TCEP 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: 6-Phosphogluconate Dehydrogenase/PGD

6-phosphogluconate dehydrogenase (PGD) is the second dehydrogenase in the pentose phosphate pathway (1). It catalyzes the oxidative decarboxylation of
6-phosphogluconate to ribulose 5-phosphate and CO2, with concomitant reduction of NADP to NADPH (2). Together with glucose-6-phosphate dehydrogenase (G6PD), these enzymes ensure the reducing environment of the cell cytoplasm (3). However, the activities of both enzymes are strongly inhibited by NADPH. It is not quite clear how the enzymes are regulated to maintain the level of reducing power inside cells (4). Deficiency of PGD is generally asymptomatic, and the inheritance of this disorder is autosomal dominant. Hemolysis results from combined deficiency of PGD and 6-phosphogluconolactonase suggest a synergism of the two enzymopathies (5).

References

  1. Barcia-Vieitez, R. and Ramos-Martinez, J.I. (2114) IUBMB Life 66:775.
  2. Wamelink, M.M. et al. (2008) J. Inherit. Metab. Dis. 31:703.
  3. Tsui, S.K.W. et al. (1996) Biochem. Genet. 34:367.
  4. Zimmer, H.G. (1996) Mol. Cell Biochem. 160-161:101.
  5. Caprari, P. et al. (2001) Ann. Hematol. 80:41.

Long Name

6-Phosphogluconate Dehydrogenase

Alternate Names

6PGD, PGDH

Entrez Gene IDs

5226 (Human); 110208 (Mouse); 362660 (Rat)

Gene Symbol

PGD

UniProt

P52209

Additional 6-Phosphogluconate Dehydrogenase/PGD Products

Product Documents for Recombinant Human 6-Phosphogluconate Dehydrogenase/PGD, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Human 6-Phosphogluconate Dehydrogenase/PGD, CF

For research use only

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Protocols

View specific protocols for Recombinant Human 6-Phosphogluconate Dehydrogenase/PGD, CF (8964-DH):

Materials
  • Assay Buffer: 25 mM MES, 150 mM NaCl, 10 mM MgCl2, pH 7.0
  • Recombinant Human 6-Phosphogluconate Dehydrogenase/PGD (rhPGD) (Catalog # 8964-DH)
  • Donor: 6-phosphogluconic acid trisodium salt, 20 mM stock in 50 mM HCl
  • Acceptor:  beta -Nicotinamide adenine dinucleotide phosphate (NADP+), 50 mM stock in deionized water
  • 96-well Clear Plate (Catalog # DY990)
  • Plate Reader
  1. Dilute rhPGD to 1 µg/mL in Assay Buffer.
  2. Prepare Reaction Mixture containing 1 mM NADP+ and 0.4 mM 6-phosphogluconic acid trisodium salt in Assay Buffer.
  3. Load 50 µL of 1 µg/mL rhPGD in a plate, and start the reaction by adding 50 µL of Reaction Mixture. Include a Control containing 50 µL of Assay Buffer and 50 µL of Reaction Mixture.
  4. Read at 340 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 Control
      **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:

  • rhPGD: 0.05 µg
  • NADP+: 0.5 mM
  • 6-phosphogluconic acid trisodium salt: 0.2 mM

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