Recombinant Human ENPP-2/Autotaxin Protein, CF

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Product Details

Summary
Reactivity HuSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant Human ENPP-2/Autotaxin Protein, CF Summary

Details of Functionality
Measured by its ability to cleave Bis (p-Nitrophenyl) Phosphate (BPNPP). The specific activity is >8,000 pmol/min/μg, as measured under the described conditions.
Source
Mouse myeloma cell line, NS0-derived human ENPP-2/Autotaxin protein
Asp49-Ile863, with an N-terminal 6-His tag
Accession #
N-terminal Sequence
His
Protein/Peptide Type
Recombinant Enzymes
Gene
ENPP2
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by silver stain
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
94 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
99 kDa and 103 kDa, migrates as a doublet under reducing conditions
Publications
Read Publications using
5255-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 silver stain
Assay Procedure
  • Assay Buffer: 50 mM Tris, 10 mM CaCl2, 5 mM MgCl2, 0.02% Brij-35 (v/v), pH 8.5
  • Recombinant Human ENPP‑2/Autotaxin (rhENPP-2) (Catalog # 5255-EN)
  • Substrate: Bis(p-Nitrophenyl) Phosphate Sodium Salt (BPNPP) (Sigma, Catalog # N3002), 40 mM stock in deionized water (Note: Heating may be necessary to solubilize Substrate.)
  • 96-well Clear Plate (Costar, Catalog # 92592)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhENPP-2 to 0.1 ng/µL in Assay Buffer.
  2. Dilute Substrate BPNPP to 2 mM in Assay Buffer.
  3. Load into the wells of a microplate 50 µL of 0.1 ng/µL rhENPP-2 and start the reaction by adding 50 µL of 2 mM Substrate. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL Substrate.
  4. Read at 400 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 p-Nitrophenol (Sigma-Aldrich, Catalog # 241326).

Per Well:
  • rhENPP-2: 0.005 µg
  • Substrate: 1 mM

Notes

This product may be covered by one or more of the following US issued patents 5,449,753; 5,731,167; 6,084,069; 6,417,338 and US pending applications.



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

Alternate Names for Recombinant Human ENPP-2/Autotaxin Protein, CF

  • ATX
  • ATXFLJ26803
  • ATX-X
  • Autotaxin
  • autotaxin-t
  • EC 3.1.4.39
  • ectonucleotide pyrophosphatase/phosphodiesterase 2
  • ectonucleotide pyrophosphatase/phosphodiesterase family member 2
  • E-NPP 2
  • ENPP2
  • ENPP-2
  • Extracellular lysophospholipase D
  • Lysophosphatidic Acid
  • LysoPLD
  • NPP2
  • PD-IALPHA
  • PDNP2
  • PDNP2NPP2
  • phosphodiesterase I/nucleotide pyrophosphatase 2
  • plasma lysophospholipase D

Background

ENPP-2, also known as Autotaxin, belongs to the ectonucleotide pyrophosphatase/phosphodiesterase (NPP) family. Some NPPs hydrolyze phosphates from nucleotides and their derivatives. ENPP-2 shares 40 - 50% identity to ENPP1 & 3, all of which contain a N-terminal intracellular domain, a single transmembrane domain and a large extracellular domain that includes a catalytic domain, two somatomedin-B-like domains, and a C-terminal nuclease-like domain (1). Unlike ENPP-1 and ENPP-3, ENPP-2 has weak activity against nucleotides, but exhibits a lysophospholipase D activity which allows the formation of lysophosphatidic acid (LPA) and choline from lysophosphatidylcholine (2). The hydrolysis of nucleotides and lysophospholipids by ENPP-2 is mediated by a single catalytic site (2). Evidence shows LPA and sphingosine 1-phosphate to be specific inhibitors of ENPP-2 (3). ENPP-2 was originally found to stimulate tumor cell motility and has since been found to enhance tumor invasion and metastasis (4) and to be up-regulated in several types of carcinomas including breast and lung (5).

  1. Cimpean, A. et al. (2004) Biochem. J. 381:71.
  2. Gijsbers, R. et al. (2003) FEBS Letters. 538:60.
  3. Van Meeteren, L.A. et al. (2005) J. Biol. Chem. 280:21155.
  4. Nam, S.W. et al. (2000) Oncogene 19:241.
  5. Jansen, S. et al. (2005) J. Cell Sci. 118:3081.

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Publications for ENPP-2/Autotaxin (5255-EN)(4)

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Bioinformatics

Gene Symbol ENPP2
Entrez
Uniprot