ENPP2 ELISA Kit (Rat)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Ectonucleotide Pyrophosphatase/Phosphodiesterase 2 (ENPP2) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Ectonucleotide Pyrophosphatase/Phosphodiesterase 2 (ENPP2) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

Autotaxin; ectonucleotide pyrophosphatase/phosphodiesterase family member 2; E-NPP 2; extracellular lysophospholipase D; lysoPLD; phosphodiesterase I/nucleotide pyrophosphatase 2 (autotaxin) .

Hydrolyzes lysophospholipids to produce the signaling molecule lysophosphatidic acid (LPA) in extracellular fluids (PubMed:12633853, PubMed:29259743, PubMed:27660691, PubMed:27268273, PubMed:27075612) . Major substrate is lysophosphatidylcholine (PubMed:12119361, PubMed:27660691, PubMed:27268273, PubMed:27075612) . Also can act on sphingosylphosphorylcholine producing sphingosine-1-phosphate, a modulator of cell motility (PubMed:12119361) . Can hydrolyze, in vitro, bis-pNPP, to some extent pNP-TMP, and barely ATP (PubMed:12633853, PubMed:27268273) . Involved in several motility-related processes such as angiogenesis and neurite outgrowth. Acts as an angiogenic factor by stimulating migration of smooth muscle cells and microtubule formation. Stimulates migration of melanoma cells, probably via a pertussis toxin-sensitive G protein. May have a role in induction of parturition. Possible involvement in cell proliferation and adipose tissue development. Tumor cell motility-stimulating factor (By similarity) .

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Ectonucleotide Pyrophosphatase/Phosphodiesterase 2 (ENPP2) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Ectonucleotide Pyrophosphatase/Phosphodiesterase 2 (ENPP2) . Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Ectonucleotide Pyrophosphatase/Phosphodiesterase 2 (ENPP2), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of Ectonucleotide Pyrophosphatase/Phosphodiesterase 2 (ENPP2) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

Matrix Recovery range (%) Average (%) Serum (n=5) 98-105 102 EDTA Plasma (n=5) 96-104 101 Heparin Plasma (n=5) 96-104 101 Matrix Recovery range (%) Average (%) Matrix Recovery range (%) Average (%) Serum (n=5) 98-105 102 Serum (n=5) 98-105 102 EDTA Plasma (n=5) 96-104 101 EDTA Plasma (n=5) 96-104 101 Heparin Plasma (n=5) 96-104 101 Heparin Plasma (n=5) 96-104 101

Component Amount Anti-ENPP2 Microplate 96 Wells (12 x 8 Well strips) ENPP2 Lyophilized Standard 2 100X Biotinylated ENPP2 Detector Antibody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL Component Amount Component Amount Anti-ENPP2 Microplate 96 Wells (12 x 8 Well strips) Anti-ENPP2 Microplate 96 Wells (12 x 8 Well strips) ENPP2 Lyophilized Standard 2 ENPP2 Lyophilized Standard 2 100X Biotinylated ENPP2 Detector Antibody 1 x 120 uL 100X Biotinylated ENPP2 Detector Antibody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL

This assay has high sensitivity and excellent specificity for detection of Ectonucleotide Pyrophosphatase/Phosphodiesterase 2 (ENPP2) . No significant cross-reactivity or interference between Ectonucleotide Pyrophosphatase/Phosphodiesterase 2 (ENPP2) and analogues was observed.