Recombinant Human ENPP-5 Protein, CF

R&D Systems | Catalog # 8655-EN

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Key Product Details

  • R&D Systems HEK293-derived Recombinant Human ENPP-5 Protein (8655-EN)
  • 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

Q9UJA9

Applications

Enzyme Activity
View all ENPP-5 Proteins and Enzymes »

Product Specifications

Source

Human embryonic kidney cell, HEK293-derived human ENPP-5 protein
Pro25-Ser430, with a C-terminal 10-His tag

Purity

>90%, 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

Pro25

Predicted Molecular Mass

48 kDa

SDS-PAGE

70-80 kDa, reducing conditions

Activity

Measured by its ability to cleave O-(4-Nitrophenylphosphoryl) choline.
The specific activity is >300 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

8655-EN
Formulation Supplied as a 0.2 μm filtered solution in Tris and NaCl.
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: ENPP-5

Ectonucleotide pyrophosphatase/phosphodiesterase 5 (ENPP-5 or NPP5) belongs to a group of ecto-enzymes which regulate the availability of extracellular nucleotides (1). This enzyme family forms a subgroup of a larger family that also includes arylsulfatases, phosphopentomutases, 2,3-bisphosphoglycerate-independent phosphoglycerate mutases (iPGM), and alkaline phosphatases (2). Mature human ENPP-5 is an approximately 50 kDa protein that consists of a 407 amino acid (aa) extracellular region with one phosphodiesterase/nucleotide pyrophosphatase domain, a 21 aa transmembrane segment, and a 25 aa cytoplasmic tail (3, 4). Within the ectodomain, human ENPP-5 shares 85% aa sequence identity with mouse and rat ENPP-5. R&D Systems in-house testing indicates that recombinant human ENPP-5 exhibits phosphodiesterase activity with the substrate O-(4-Nitrophenylphosphoryl) choline.

References

  1. Zimmermann, H. et al. (2012) Purinergic Signal. 8:437.
  2. Gijsbers, R. et al. (2001) J. Biol. Chem. 276:1361.
  3. Ohe, Y. et al. (2003) Biochem. Biophys. Res. Commun. 308:719.

Long Name

Ectonucleotide Pyrophosphatase/Phosphodiesterase 5

Alternate Names

ENPP5, NPP5

Entrez Gene IDs

59084 (Human); 83965 (Mouse)

Gene Symbol

ENPP5

UniProt

Q9UJA9

Additional ENPP-5 Products

Product Documents for Recombinant Human ENPP-5 Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Human ENPP-5 Protein, CF

For research use only

Citations for Recombinant Human ENPP-5 Protein, CF

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Protocols

View specific protocols for Recombinant Human ENPP-5 Protein, CF (8655-EN):

Materials
  • Assay Buffer: 50 mM Sodium Acetate, 150 mM NaCl, pH 5.5
  • Recombinant Human ENPP-5 (rhENPP-5) (Catalog # 8655-EN)
  • Substrate: O-(4-Nitrophenylphosphoryl) choline (Sigma, Catalog # N5879), 500 mM stock in deionized water
  • NaOH, 0.2 M in deionized water
  • 96-well Clear Plate (Catalog # DY990)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhENPP-5 to 5 µg/mL in Assay Buffer.
  2. Dilute Substrate to 2 mM in Assay Buffer.
  3. Load 50 µL of 5 µg/mL rhENPP-5 in a clear strip well plate, and start the reaction by adding 50 µL of 2 mM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL 2 mM Substrate.
  4. Incubate sealed plate at room temperature for 30 minutes.
  5. Stop reactions by adding 100 µL of 0.2 M NaOH to each well used.
  6. Read at 410 nm (absorbance) in endpoint mode.
  7. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

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

    
      *Adjusted for Substrate Blank
      **Derived using calibration standard p-Nitrophenol (Sigma, Catalog # 241326).

Per Well:

  • rhENPP-5: 0.25 µg
  • Substrate: 1 mM

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