Recombinant Human HPGDS Protein, CF

R&D Systems | Catalog # 6487-PS

R&D Systems
Discontinued Product
6487-PS has been discontinued. View all Hematopoietic Prostaglandin D Synthase/HPGDS products.

Key Product Details

  • R&D Systems E. coli-derived Recombinant Human HPGDS Protein (6487-PS)
  • 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

Applications

Enzyme Activity
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Product Specifications

Source

E. coli-derived human Hematopoietic Prostaglandin D Synthase/HPGDS protein
Met1-Leu199, with a C-terminal 6-His tag

Purity

>95%, 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

Pro2

Predicted Molecular Mass

24 kDa

SDS-PAGE

24-26 kDa, reducing conditions

Activity

Measured by the conjugation of reduced glutathione to 1-bromo-2,4-dinitrobenzene.
The specific activity is >650 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

6487-PS
Formulation Supplied as a 0.2 μm filtered solution in Tris and NaCl.
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: Hematopoietic Prostaglandin D Synthase/HPGDS

Prostaglandin D Synthase (PGDS) catalyzes the conversion of prostaglandin (PG) H2 to PGD2, which is a major prostanoid produced in a variety of tissues. Two types of PGDS have been isolated; the glutathione-dependent hematopoietic PGDS (HPGDS) and the glutathione-independent lipocalin‑type PGDS (1). HPGDS is a cytosolic enzyme that is expressed in mast cells and antigen presenting cells (2, 3). It is the only mammalian member of the class Sigma glutathione S‑transferase, showing a broad specificity towards standard transferase substrates (4). The PGD2 produced by HPGDS is involved in many physiological processes such as maintaining body temperature, promotion of sleep, inhibition of platelet aggregation and bronchoconstriction (5). It also functions in immune response and acts as a mediator in allergy and inflammation (6). HPGDS‑specific inhibitors may be therapeutically useful anti‑allergic and anti‑inflammatory drugs.

References

  1. Urade, Y. and Eguchi, N. (2002) Prostaglandins Other Lipid Mediat. 68:375.
  2. Urade, Y. et al. (1990) J. Biol. Chem. 265:371.
  3. Urade, Y. et al. (1989) J. Immunol. 143:2982.
  4. Jowsey, I. R. et al. (2001) Biochem. J. 359:507.
  5. Kanaoka, Y. and Urade, Y. (2003) Prostaglandins Leukot. Essent. Fatty Acids 69:163.
  6. Oguma, T. et al. (2008) Allergol. Int. 57:307.

Alternate Names

GSTS, PGDS, PTGDS2

Entrez Gene IDs

27306 (Human); 54486 (Mouse); 58962 (Rat)

Gene Symbol

HPGDS

UniProt

Additional Hematopoietic Prostaglandin D Synthase/HPGDS Products

Product Documents for Recombinant Human HPGDS 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.

Product Specific Notices for Recombinant Human HPGDS Protein, CF

Coomassie is a registered trademark of Imperial Chemical Industries Ltd.

For research use only

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Protocols

View specific protocols for Recombinant Human HPGDS Protein, CF (6487-PS):

Materials
  • Assay Buffer: 100 mM NaH2PO4, pH 7.0
  • Recombinant Human Hematopoietic Prostaglandin D Synthase/HPGDS (rhHPGDS) (Catalog # 6487-PS)
  • Substrate: 1-bromo-2,4-dinitrobenzene (BDNB) (Sigma, Catalog # 262226), 75 mM stock in ethanol
  • L-Glutathione, reduced (GSH) (Amresco, Catalog # 399), 250 mM stock in deionized water
  • UV Plate (Costar, Catalog # 3635)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhHPGDS to 40 ng/μL in Assay Buffer.
  2. Dilute the GSH to 4 mM in Assay Buffer.
  3. Combine equal volumes of 40 ng/μL rhHPGDS and 4 mM GSH for 20 ng/μL rhHPGDS with 2 mM GSH.
  4. Dilute Substrate to 2 mM in Assay Buffer.
  5. Load into a UV plate 50 μL of the 20 ng/μL rhHPGDS with 2 mM GSH mixture. Include a substrate blank containing 25 μL of Assay Buffer with 25 μL of the 4 mM GSH prepared in step 2.
  6. Start the reaction by adding 50 μL of 2 mM Substrate to wells.
  7. Read in kinetic mode for 5 minutes at an absorbance of 340 nm.
  8. 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 9600 M-1cm-1 
     ***Using the path correction 0.320 cm
     Note: the output of many spectrophotometers is in mOD. Per Well:
  • rhHPGDS: 1 μg
  • GSH: 1 mM
  • Substrate: 1 mM

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