Recombinant Human COX-2 His-tag Protein, CF

R&D Systems | Catalog # 10465-CX

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Key Product Details

  • R&D Systems Sf 21 (baculovirus)-derived Recombinant Human COX-2 His-tag Protein (10465-CX)
  • 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

Sf 21 (baculovirus)

Accession Number

P35354.2

Applications

Enzyme Activity
View all COX-2 Proteins and Enzymes »

Product Specifications

Source

Spodoptera frugiperda, Sf 21 (baculovirus)-derived human COX-2 protein
Ala18-Leu604
with a C-terminal 6-His tag

Purity

>90%, 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

Ala18

Predicted Molecular Mass

68 kDa

SDS-PAGE

65-71 kDa, under reducing conditions.

Activity

Measured by its ability to convert arachidonic acid to prostaglandin H2.

Scientific Data Images for Recombinant Human COX-2 His-tag Protein, CF

Recombinant Human COX-2 His-tag Protein SDS-PAGE.

2 μg/lane of Recombinant Human COX-2 His-tag Protein (Catalog # 10465-CX) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 65-71 kDa.

Formulation, Preparation, and Storage

10465-CX
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl, CHAPS 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: COX-2

Cyclooxygenase 2 (COX-2), also known as Prostaglandin synthase 2 (PTGS2), is a dimeric, heme-dependent enzyme where the dimer is formed from two monomers each containing an N-terminal EGF-like domain, a membrane binding domain and a C-terminal catalytic domain (1). COX-2 catalyzes the conversion of arachidonate to prostaglandin H2 through a two-step reaction, representing the rate-limiting enzyme in the biosynthesis of prostanoids. Altered levels of prostaglandins are associated with several disease pathologies.  There are two major isozymes of PTGS with slightly differing substrate preferences; PTGS1 represents the constitutively expressed form while PTGS2 basal expression is present in a limited number of tissues (2) such as inflammatory cells. PTGS2 is upregulated in chronic and acute inflammation as well as in most types of cancers (3) resulting in increased rate of recurrence (4), reduction in survival (5), and increased resistance to chemo and radiotherapy (6) through the role it plays in multiple pathways and mechanisms (7). Inhibition of PTGS2 is a promising therapeutic target to reduce cancer risk (8, 9). COX-2 is a pharmacological target of nonsteroidal anti-inflammatory drugs (NSAIDS) and COX-2 selective inhibitors (coxibs) as well as other categories of inhibitors through multiple mechanisms of action (1). It remains an attractive target for novel specific inhibitor development as inhibition shows equivalent efficacy to use of conventional NSAIDs, but with reduced negative side effects (10, 11).

References

  1. Orlando, B.J. et al. (2015) J. Struct. Biol. 189:62.
  2. Su, C.W. et al. (2016) Crit. Rev. Oncol. Hematol. 107:33.
  3. Gurram, B. et al. (2018) Anal. Chem. 90:5187.
  4. Montezuma, M.A.P. et al. (2018) Pathol. Res. Pract. 214:907.
  5. Hoing, B. et al. (2018) Oncotarget 9:8415.
  6. Dong, X.F. et al. (2018) Clin. Cancer Res. 24:3204.
  7. Hashemi Goradel, N. et al. (2019) J. Cell Physiol. 234:5683.
  8. Brizzolara, A. et al. (2017) Cancer Letters 400:9.
  9. Shi, L. et al. (2018) ACS Appl. Mater. Interfaces 10:8555.
  10. Ferrer, M.D. et al. (2019) Curr. Med. Chem. 26:3225.
  11. Carullo, G. et al. (2016) Medchemcomm. 8:492.

Long Name

Cyclooxygenase 2

Alternate Names

COX2, PGHS-2, PHS-II, PTGS2

Entrez Gene IDs

5743 (Human); 19225 (Mouse); 29527 (Rat)

Gene Symbol

PTGS2

UniProt

P35354.2

Additional COX-2 Products

Product Documents for Recombinant Human COX-2 His-tag Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Human COX-2 His-tag Protein, CF

For research use only

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Protocols

View specific protocols for Recombinant Human COX-2 His-tag Protein, CF (10465-CX):

Materials
  •  Assay Buffer A: 50 mM Tris, 5 µM Hemin (Sigma, Catalog # H9039), pH 8.0
  •  Assay Buffer B: 50 mM Tris, pH 8.0
  •  Recombinant Human COX-2 (rhCOX-2) (Catalog # 10465-CX)
  •  Substrate Component 1: Arachidonic acid (Sigma, Catalog # 10931), 10 mM stock in DMSO
  •  Substrate Component 2: Amplex Ultra Red (AUR) (Molecular Probes, Catalog # A36006), 10 mM stock in DMSO
  •  F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  •  Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rhCOX-2 to 5 ng/µL in Assay Buffer A.
  2. Prepare Substrate Mixture 50 µM Arachidonic acid and 100 µM AUR in Assay Buffer B.
  3. Load into a plate 50 µL of diluted rhCOX-2.  Also create a Substrate Blank by loading 50 µL of Assay Buffer A.
  4. Add 50 µL of Substrate Mixture to all wells and incubate at room temperature for 1 minute.
  5. Read at excitation and emission wavelengths of 540 nm and 585 nm (top read), respectively in endpoint mode.
  6. Calculate specific activity:
 

     Specific Activity (pmol/min/µg) =

Adjusted Fluorescence* (RFU) x Conversion Factor** (pmol/RFU)
Incubation time (min) x amount of enzyme (µg)

*Adjusted for Substrate Blank
**Derived using calibration standard Resorufin (Sigma, Catalog # R3257)

Per Well
  • rhCOX-2: 0.25 µg
  • Arachidonic acid: 25 µM
  • AUR: 50 µM

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