Recombinant Human PON1 His-tag Fc Chimera Protein, CF

R&D Systems | Catalog # 10175-PO

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Key Product Details

  • R&D Systems HEK293-derived Recombinant Human PON1 His-tag Fc Chimera Protein (10175-PO)
  • 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

HEK293

Accession Number

P27169

Applications

Enzyme Activity
View all PON1 Proteins and Enzymes »

Product Specifications

Source

Human embryonic kidney cell, HEK293-derived human PON1 protein
Leu16-Leu355
with an N-terminal 6-His tag and a C-terminal Fc 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

His

Predicted Molecular Mass

66 kDa

SDS-PAGE

69-79 kDa, under reducing conditions

Activity

Measured by its ability to hydrolyze phenyl acetate.
The specific activity is >5000 pmol/min/μg, as measured under the described conditions.

Scientific Data Images for Recombinant Human PON1 His-tag Fc Chimera Protein, CF

Recombinant Human PON1 His-tag Fc Chimera Protein Enzyme Activity

Recombinant Human PON1 His-tag Fc Chimera Protein Enzyme Activity

Recombinant Human PON-1 His-tag Fc Chimera (Catalog # 10175-PO) is measured by its ability to hydrolyze phenyl acetate. The activity (orange) is over 3-fold higher than the competitor's PON-1 (green).
Recombinant Human PON1 His-tag Fc Chimera Protein SDS-PAGE

Recombinant Human PON1 His-tag Fc Chimera Protein SDS-PAGE

2 μg/lane of Recombinant Human PON1 His-tag Fc Chimera was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® blue staining, showing a band at ~74 kDa under reducing conditions.

Formulation, Preparation, and Storage

10175-PO
Formulation Supplied as a 0.2 μm filtered solution in Tris, CaCl2, NaCl, 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: PON1

Serum paraoxonase 1 (PON-1) is a member of the paraoxonase family which has three members: PON1, PON2, PON3.  PON-1 is a lactonase (1,2) but has significant promiscuity (2,3) that allows hydrolysis of a variety of substrates including organophosphate triesters (including many pesticides), arylesters, cyclic carbamates, glucuronides and consequently also pharmaceuticals such as statins. PON-1 is a calcium-dependent, secreted, 43 kDa protein with a 6-bladed propeller structure and an active site gorge containing a His-His catalytic dyad (4).  It is a homodimer and retains its hydrophobic signal sequence in the N-terminal region which enables its association with HDL, resulting in its localization predominantly in the plasma (5,6). Low serum PON1 and dysfunctional HDL is associated with many diseases with an inflammatory component including diabetes mellitus (7,8), rheumatoid arthritis, hepatic and renal diseases, psoriasis, and macular degeneration (9,10). PON1 shows a variety of atheroprotective properties (6,11,12) by metabolizing inflammatory lipid peroxides (13). PON-1 activity is low in coronary heart disease patients (14) and contributes to an increased risk of a major adverse cardiovascular event (15). PON-1 also protects against toxicity of organophosphorus compounds used as pesticides (16,17) thought to increase risk of neurodegenerative disorders such as Parkinson's (18) and Amyotrophic Lateral Sclerosis (19).

References

  1. Dragonov, D. I. et al. (2005) Lipid Res. 46:1239.
  2. Khersonsky, O. et al. (2005) Biochemistry. 44:6371.
  3. Ben-David, M. et al. (2012) J. Mol. Biol. 418:181.
  4. Harel, M. et al. (2004) Nature Struct. Mol. Biol. 11:412.
  5. Mackness, B. et al. (1998) Gen. Pharmac. 31:329.
  6. Aviram, M. et al. (2004) Free Radic. Biol. Med. 37:1304.
  7. Rozenberg, O. et al. (2008) Free Radic. Biol. Med. 44:1951.
  8. Koren-Gluzer, M. et al. (2011) Atherosclerosis. 219:532.
  9. Goswami, N. et al. (2009) Clin. Chim. Acta. 410:1.
  10. Mahrooz, A. (2016) Curr. Clin. Pharmacol. 11:259.
  11. Berrougui, H. et al. (2012) Free Radic. Biol. Med. 52:1372.
  12. Aharoni, S. et al. (2013) Atherosclerosis 228:353.
  13. Tavori, H. (2011) Free Rad. Biol. Med. 51:234.
  14. Wang, M. et al. (2012) DNA Cell. Biol. 31:975.
  15. Tang, W. H. W. et al. (2012) Arterioscler. Thromb. Vasc. Biol. 32:2803.
  16. Li, W. F. et al. (2000) Pharmacogenetics. 10:767.
  17. Shih, D. M. et al. (1998) Nature. 394:284.
  18. Ahmed, H. et al. (2017) Biomed. Pharmacother. 90:638.
  19. Gagliardi, S. et al. (2013) Neurotox. Res. 23:370.

Long Name

Paraoxonase 1

Alternate Names

ESA, K-45

Entrez Gene IDs

5444 (Human); 18979 (Mouse); 84024 (Rat)

Gene Symbol

PON1

UniProt

P27169 (Human)

Additional PON1 Products

Product Documents for Recombinant Human PON1 His-tag Fc Chimera Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Human PON1 His-tag Fc Chimera Protein, CF

For research use only

Related Research Areas

Cancer Stem Cell Markers

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Protocols

View specific protocols for Recombinant Human PON1 His-tag Fc Chimera Protein, CF (10175-PO):

Materials
  • Assay Buffer: 50 mM Tris, 5 mM CaCl2, pH 8.0
  • Deionized Water
  • Recombinant Human PON1 His-tag Fc Chimera (rhPON1) (Catalog 10175-PO)
  • Substrate: Phenyl Acetate (7.88 M) (Sigma, Catalog # 108723)
  • UV Plate (Catalog # 3635)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhPON1 to 10 µg/mL in Assay Buffer.
  2. Dilute Phenyl Acetate to 40 mM in deionized water.
  3. Load 50 µL of 10 µg/mL rhPON1 in a plate, and start the reaction by adding 50 µL of Substrate.  Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of Substrate.
  4. Read plate at a wavelength of 270 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 1310 M-1cm-1
***Using the path correction 0.32 cm
Note: the output of many spectrophotometers is in mOD

Per Well:

  • rhPON1: 0.5 µg
  • Phenyl Acetate: 20 mM

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