< 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.
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
The recovery of ENPP-2 spiked to levels throughout the range of the assay was evaluated.
Average % Recovery
Cell Culture Media (n=4)
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.
Preparation and Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
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.
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.
Prepare all reagents, standard dilutions, and samples as directed in the product insert.
Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
100 µL Assay Diluent
Add 100 µL of Assay Diluent to each well.
50 µL Standard, Control, or Sample
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.
Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
200 µL Conjugate
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.
Aspirate and wash 4 times.
200 µL Substrate Solution
Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
50 µL Stop Solution
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.
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The data collected includes not only links to publications in PubMed,
but also provides information about sample types, species, and experimental conditions.