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Recombinant Human ENPP-5 Protein, CF

Catalog #: 8655-EN
1 Citation Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
8655-EN-020
R&D Systems Recombinant Proteins and Enzymes
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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human ENPP-5 Protein, CF Summary

Product Specifications

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.
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.
Source
Human embryonic kidney cell, HEK293-derived human ENPP-5 protein
Pro25-Ser430, with a C-terminal 10-His tag
Accession #
Q9UJA9
N-terminal Sequence
Analysis
Pro25
Predicted Molecular Mass
48 kDa
SDS-PAGE
70-80 kDa, reducing conditions

Product Datasheets

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8655-EN

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.

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.

Assay Procedure

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
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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
Entrez Gene IDs
59084 (Human); 83965 (Mouse)
Alternate Names
EC 3.1; ectonucleotide pyrophosphatase/phosphodiesterase 5 (putative function); ectonucleotide pyrophosphatase/phosphodiesterase 5 (putative); ectonucleotide pyrophosphatase/phosphodiesterase family member 5; E-NPP 5; ENPP5; ENPP-5; KIAA0879; NPP5; NPP-5

Citation for Recombinant Human ENPP-5 Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

  1. Identification of the extracellular membrane protein ENPP3 as a major cGAMP hydrolase and innate immune checkpoint
    Authors: Mardjuki, R;Wang, S;Carozza, J;Zirak, B;Subramanyam, V;Abhiraman, G;Lyu, X;Goodarzi, H;Li, L;
    Cell reports
    Species: N/A
    Sample Types: Small Molecule
    Applications: Bioassay

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