Recombinant Human ENPP-2/Autotaxin Protein, CF

Catalog # Availability Size / Price Qty
5255-EN-010
Product Details
Citations (5)
FAQs
Reviews

Recombinant Human ENPP-2/Autotaxin Protein, CF Summary

Product Specifications

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.
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.
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
Analysis
His
Predicted Molecular Mass
94 kDa
SDS-PAGE
99 kDa and 103 kDa, migrates as a doublet under reducing conditions

Product Datasheets

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.

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.
  • 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 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
  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
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-2/Autotaxin

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).

References
  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.
Long Name
Ectonucleotide Pyrophosphatase/Phosphodiesterase 2
Entrez Gene IDs
5168 (Human); 18606 (Mouse); 84050 (Rat)
Alternate Names
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

Citations for Recombinant Human ENPP-2/Autotaxin 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.

5 Citations: Showing 1 - 5
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  1. The involvement of autotaxin in renal interstitial fibrosis through regulation of fibroblast functions and induction of vascular leakage
    Authors: N Sakai, G Bain, K Furuichi, Y Iwata, M Nakamura, A Hara, S Kitajima, A Sagara, T Miyake, T Toyama, K Sato, S Nakagawa, M Shimizu, S Kaneko, T Wada
    Sci Rep, 2019;9(1):7414.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  2. Activation of Adenylyl Cyclase Causes Stimulation of Adenosine Receptors
    Authors: T Pleli, A Mondorf, N Ferreiros, D Thomas, K Dvorak, RM Biondi, DMZ Heringdorf, S Zeuzem, G Geisslinge, H Zimmermann, O Waidmann, A Piiper
    Cell. Physiol. Biochem., 2018;45(6):2516-2528.
    Species: N/A
    Sample Types: Small Molecule
    Applications: Bioassay
  3. Proteomics Based Identification of Autotaxin As An Anti-Hepatitis B Virus Factor and a Promoter of Hepatoma Cell Invasion and Migration
    Authors: S She, M Yang, H Hu, P Hu, Y Yang, H Ren
    Cell. Physiol. Biochem., 2018;45(2):744-760.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  4. ATX?LPA axis facilitates estrogen?induced endometrial cancer cell proliferation via MAPK/ERK signaling pathway
    Authors: G Zhang, Y Cheng, Q Zhang, X Li, J Zhou, J Wang, L Wei
    Mol Med Rep, 2018;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay
  5. Autotaxin-LPA axis regulates hMSC migration by adherent junction disruption and cytoskeletal rearrangement via LPAR1/3-dependent PKC/GSK3beta/beta-catenin and PKC/Rho GTPase pathways.
    Authors: Ryu J, Han H
    Stem Cells, 2015;33(3):819-32.
    Species: Human
    Sample Types: Whole Cells
    Applications: Bioassay

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