Recombinant Human ENPP-1 Protein, CF

R&D Systems | Catalog # 6136-EN

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Key Product Details

  • R&D Systems NS0-derived Recombinant Human ENPP-1 Protein (6136-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

NS0

Accession Number

P22413 (ENPP-1) & Q13822 (ENPP-2)

Applications

Enzyme Activity
View all ENPP-1 Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived human ENPP-1 protein

His ENPP-2
Asp49-Trp144
Accession # Q13822		 ENPP-1
Val191-Leu591
  Accession # P22413 		 ENPP-2
Asn532-Ile863
Accession # Q13822
N-terminus C-terminus

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

96 kDa

SDS-PAGE

100-135 kDa, reducing conditions

Activity

Measured by its ability to hydrolyze thymidine 5'-monophosphate p-nitrophenyl ester.
The specific activity is >35,000 pmol/min/μg, as measured under the described conditions.

Reviewed Applications

Read 1 review rated 5 using 6136-EN in the following applications:

Formulation, Preparation, and Storage

6136-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-1

Ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP-1) is a transmembrane glycoprotein that hydrolyzes nucleotides and nucleotide derivatives with the formation of nucleotide-5’-monophosphates. It is inserted into the plasma membrane by an N‑terminal transmembrane domain. Human ENPP-1 has a small N-terminal cytoplasmic domain and a large C-terminal region containing two somatomedin B-like domains, a catalytic domain and a nuclease-like domain in the extracellular space (1). Defects in the ENPP-1 gene cause arterial calcification and bone mineralization abnormalities (2). ENPP-1 polymorphism or overexpression is also associated with obesity, type II diabetes and insulin resistance, which makes modulation of ENPP-1 activity one of the targets to treat insulin resistance and related diseases (1). The recombinant hybrid enzyme recombinant human ENPP-1 consists of N-terminal somatomedin B-like domains of ENPP-2, the central catalytic phosphodiesterase domain of ENPP-1 and the C‑terminal nuclease-like domain of ENPP-2. It has been reported that this hybrid construct generates an enzyme that has higher levels of ENPP-1 activity than the wild type ENPP-1 and is specific for the ENPP-1 substrate (3).

References

  1. Goldfine, I.D. et al. (2010) Endocrine Reviews. 29:62.
  2. Hessle, L. et al. (2002) Proc. Natl. Acad. Sci. 99:9445.
  3. Anisoara, C. et al. (2004) Biochem. J. 381:71.

Long Name

Ectonucleotide Pyrophosphatase/Phosphodiesterase 1

Alternate Names

ENPP1, Ly-41 Antigen, M6S1, NPP1, NPPS, PCA1, PDNP1

Entrez Gene IDs

5167 (Human); 18605 (Mouse)

Gene Symbol

ENPP1

UniProt

P22413 (ENPP-1) & Q13822 (ENPP-2)

Additional ENPP-1 Products

Product Documents for Recombinant Human ENPP-1 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-1 Protein, CF

For research use only

Citations for Recombinant Human ENPP-1 Protein, CF

Customer Reviews for Recombinant Human ENPP-1 Protein, CF (1)

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    Name: Alexis Weston
    Application: Enzymatic activity in vitro
    Verified Customer | Posted 12/06/2017
    Recombinant Human ENPP-1 Protein, CF 6136-EN

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Protocols

View specific protocols for Recombinant Human ENPP-1 Protein, CF (6136-EN):

Materials
  • Assay Buffer: 50 mM Tris, 250 mM NaCl, pH 9.5
  • Recombinant Human ENPP-1 (rhENPP-1) (Catalog # 6136-EN)
  • Substrate: Thymidine 5’-monophosphate p-nitrophenyl ester (Sigma, Catalog # T4510), 100 mM stock in deionized water
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhENPP-1 to 1 ng/µL in Assay Buffer.
  2. Dilute Substrate to 10 mM in Assay Buffer.
  3. Load into a plate 50 µL of 1 ng/µL rhENPP-1, and start the reaction by adding 50 µL of 10 mM Substrate. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of 10 mM Substrate.
  4. Read at 405 nm (absorbance) in kinetic mode for 5 minutes.
  5. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (OD/min) x Conversion Factor** (pmol/OD)
amount of enzyme (µg)

     *Adjusted for Substrate Blank
     **Derived using calibration standard 4-Nitrophenol (Sigma-Aldrich, Catalog # 241326).

Per Well:
  • rhENPP-1: 0.05 µg
  • Substrate: 5 mM

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