Human ENPP-2/Autotaxin Quantikine ELISA Kit

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Human ENPP-2/Autotaxin Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (10 uL), Heparin Plasma (10 uL), Urine (50 uL), Human Milk (50 uL)
0.157 ng/mL
Assay Range
0.8 - 50 ng/mL (Cell Culture Supernates, Serum, Heparin Plasma, Urine, Human Milk)
Natural and recombinant human ENPP-2
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
No significant interference observed with available related molecules.

Product Summary

The Quantikine Human ENPP-2 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human ENPP-2 in cell culture supernates, serum, plasma, urine, and human milk. It contains NS0-expressed recombinant human ENPP-2 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human ENPP-2 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human ENPP-2.


Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision

Cell Culture Supernates, Serum, Heparin Plasma, Urine, Human Milk

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 6.5 13.8 28.9 6.8 14 28.9
Standard Deviation 0.18 0.51 0.74 0.32 0.55 0.83
CV% 2.8 3.7 2.6 4.7 3.9 2.9


The recovery of ENPP-2 spiked to levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 102 93-109


To assess the linearity of the assay, samples containing and/or spiked with high concentrations of ENPP-2 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human ENPP-2/Autotaxin ELISA Linearity

Scientific Data

Human ENPP-2/Autotaxin ELISA Standard Curve

Product Datasheets

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Preparation and Storage

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

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. 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. The hydrolysis of nucleotides and lysophospholipids by ENPP-2 is mediated by a single catalytic site. Evidence shows LPA and sphingosine 1-phosphate to be specific inhibitors of ENPP-2. ENPP-2 was originally found to stimulate tumor cell motility and has since been found to enhance tumor invasion and metastasis and to be up-regulated in several types of carcinomas including breast and lung.

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; 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
&#9888; WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 100 µL Assay Diluent
  4.   Add 100 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Human ENPP-2/Autotaxin Quantikine ELISA Kit

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. Autotaxin secretion is a stromal mechanism of adaptive resistance to TGF? inhibition in pancreatic ductal adenocarcinoma
    Authors: Pietrobono, S;Sabbadini, F;Bertolini, M;Mangiameli, D;De Vita, V;Fazzini, F;Lunardi, G;Casalino, S;Scarlato, E;Merz, V;Zecchetto, C;Quinzii, A;Di Conza, G;Lahn, M;Melisi, D;
    Cancer research
    Species: Human
    Sample Types: Plasma
  2. Immunometabolic analysis shows a distinct cyto-metabotype in Covid-19 compared to sepsis from other causes
    Authors: FM Trovato, S Mujib, E Jerome, A Cavazza, P Morgan, J Smith, MT Depante, K O'Reilly, J Luxton, T Mare, S Napoli, MJ McPhail
    Heliyon, 2022-06-24;8(6):e09733.
    Species: Human
    Sample Types: Plasma
  3. Serum autotaxin is a parameter for the severity of liver cirrhosis and overall survival in patients with liver cirrhosis--a prospective cohort study.
    Authors: Pleli T, Martin D, Kronenberger B, Brunner F, Koberle V, Grammatikos G, Farnik H, Martinez Y, Finkelmeier F, Labocha S, Ferreiros N, Zeuzem S, Piiper A, Waidmann O
    PLoS ONE, 2014-07-25;9(7):e103532.
    Species: Human
    Sample Types: Serum
  4. Serum autotaxin levels correlate with pruritus in patients with atopic dermatitis.
    Authors: Nakao, Momoko, Sugaya, Makoto, Suga, Hiraku, Kawaguchi, Makiko, Morimura, Sohshi, Kai, Hiromich, Ohmatsu, Hanako, Fujita, Hideki, Asano, Yoshihid, Tada, Yayoi, Kadono, Takafumi, Sato, Shinichi
    J Invest Dermatol, 2014-01-17;134(6):1745-7.
    Species: Human
    Sample Types: Serum
  5. Non-invasive imaging of tumors by monitoring autotaxin activity using an enzyme-activated near-infrared fluorogenic substrate.
    Authors: Madan, Damian, Ferguson, Colin G, Lee, Won Yong, Prestwich, Glenn D, Testa, Charles
    PLoS ONE, 2013-11-20;8(11):e79065.
    Species: Human
    Sample Types: Tissue Homogenates


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Human ENPP-2/Autotaxin Quantikine ELISA Kit
By Anonymous on 01/17/2020
Sample Tested: C6 rat glioma cell line