Recombinant Human ENPP-1 Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Human ENPP-1 Protein, CF Summary

Details of Functionality
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.
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

Accession #
N-terminal Sequence
His
Protein/Peptide Type
Recombinant Enzymes
Gene
ENPP1
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
96 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
100-135 kDa, reducing conditions
Publications
Read Publication using
6136-EN in the following applications:

Packaging, Storage & Formulations

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.
Buffer
Supplied as a 0.2 μm filtered solution in Tris and NaCl.
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane
Assay Procedure
  • 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

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human ENPP-1 Protein, CF

  • ectonucleotide pyrophosphatase/phosphodiesterase 1
  • E-NPP 1
  • ENPP1
  • ENPP-1
  • Ly-41 Antigen
  • M6S1
  • M6S1NPP1
  • Membrane component chromosome 6 surface marker 1
  • membrane component, chromosome 6, surface marker 1
  • NPP1
  • NPPS
  • NPPSalkaline phosphodiesterase 1
  • PC1
  • PC-1
  • PCA1
  • PCA1ARHR2
  • PDNP1
  • PDNP1ectonucleotide pyrophosphatase/phosphodiesterase family member 1
  • Phosphodiesterase I/nucleotide pyrophosphatase 1
  • plasma-cell membrane glycoprotein 1
  • Plasma-cell membrane glycoprotein PC-1

Background

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).
  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.

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Publications for ENPP-1 (6136-EN)(1)

We have publications tested in 1 confirmed species: N/A.

We have publications tested in 1 application: Bioassay.


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Bioinformatics