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).
Recombinant Human ENPP-2/Autotaxin Protein, CF
R&D Systems | Catalog # 5255-EN
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Key Product Details
- R&D Systems NS0-derived Recombinant Human ENPP-2/Autotaxin Protein (5255-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
Applications
Enzyme Activity
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Product Specifications
Source
Mouse myeloma cell line, NS0-derived human ENPP-2/Autotaxin protein
Asp49-Ile863, with an N-terminal 6-His tag
Asp49-Ile863, with an N-terminal 6-His tag
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
94 kDa
SDS-PAGE
99 kDa and 103 kDa, migrates as a doublet under reducing conditions
Activity
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.
The specific activity is >8,000 pmol/min/μg, as measured under the described conditions.
Reviewed Applications
Read 1 review rated 5 using 5255-EN in the following applications:
Formulation, Preparation, and Storage
5255-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.
|
Background: ENPP-2/Autotaxin
References
- Cimpean, A. et al. (2004) Biochem. J. 381:71.
- Gijsbers, R. et al. (2003) FEBS Letters. 538:60.
- Van Meeteren, L.A. et al. (2005) J. Biol. Chem. 280:21155.
- Nam, S.W. et al. (2000) Oncogene 19:241.
- Jansen, S. et al. (2005) J. Cell Sci. 118:3081.
Long Name
Ectonucleotide Pyrophosphatase/Phosphodiesterase 2
Alternate Names
ATX, Autotaxin, ENPP2, Lysophosphatidic Acid, NPP2, PDNP2
Gene Symbol
ENPP2
UniProt
Additional ENPP-2/Autotaxin Products
Product Documents for Recombinant Human ENPP-2/Autotaxin Protein, CF
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant Human ENPP-2/Autotaxin Protein, CF
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.
For research use only
Related Research Areas
Citations for Recombinant Human ENPP-2/Autotaxin Protein, CF
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Protocols
View specific protocols for Recombinant Human ENPP-2/Autotaxin Protein, CF (5255-EN):
Materials
- Assay Buffer: 50 mM Tris, 10 mM CaCl2, 5 mM MgCl2, 0.02% Brij-35 (w/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
- Dilute rhENPP-2 to 0.1 ng/µL in Assay Buffer.
- Dilute Substrate BPNPP to 2 mM in Assay Buffer.
- 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.
- Read at 400 nm (absorbance) in kinetic mode for 5 minutes.
- 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
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