Recombinant Human HPGDS Protein, CF

Catalog #: 6487-PS Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
6487-PS-010

R&D Systems Recombinant Proteins and Enzymes

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All Hematopoietic Prostaglandin D Synthase/HPGDS Proteins and Enzymes

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human HPGDS Protein, CF Summary

Product Specifications

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.

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.

Source

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

Accession #

O60760

N-terminal Sequence
Analysis

Pro2

Predicted Molecular Mass

24 kDa

SDS-PAGE

24-26 kDa, reducing conditions

Product Datasheets

6487-PS

Product Datasheet

COA

SDS

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.

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.

Assay Procedure

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

Dilute rhHPGDS to 40 ng/μL in Assay Buffer.
Dilute the GSH to 4 mM in Assay Buffer.
Combine equal volumes of 40 ng/μL rhHPGDS and 4 mM GSH for 20 ng/μL rhHPGDS with 2 mM GSH.
Dilute Substrate to 2 mM in Assay Buffer.
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.
Start the reaction by adding 50 μL of 2 mM Substrate to wells.
Read in kinetic mode for 5 minutes at an absorbance of 340 nm.
Calculate specific activity:

Specific Activity (pmol/min/µg) =	Adjusted V_max* (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

Reconstitution Calculator

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

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

Entrez Gene IDs

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

Alternate Names

EC 2.5.1.18; EC 5.3.99.2; glutathione S-transferase sigma; Glutathione S-transferase; Glutathione-dependent PGD synthase; Glutathione-requiring prostaglandin D synthase; GST class-sigma; GSTS; GSTShematopoietic prostaglandin D2 synthase; hematopoietic prostaglandin D synthase; HPGDS; H-PGDS; PGDS; PGDSglutathione-dependent PGD synthetase; Prostaglandin-H2 D-isomerase; PTGDS2