Autotaxin ELISA Kit (ATX)

Full Name: Autotaxin ELISA Kit (ATX)
Reactivity: Human
Sample Type: Plasma, Serum
Sensitivity: 0.78 ng/ml

INTRODUCTION

Autotaxin (ATX) is classified as a member of the ectonucleotide pyrophosphatase and phosphodiesterase (which can also be abbreviated as ENPP) family. It was initially identified in tumor motility stimulating proteins. It exists as a homodimer with two subunits: a large subunit (which contains its active site) and a small subunit. It is a Ca-dependent enzyme, the catalytic domain has the four conserved histidines (D123, D124, D125 and D126) which are important for catalysis. It also has an active site containing serine residue 684 and tryptophan 627 that coordinate with a water molecule in its substrate acetylcholine to cleave it into choline and acetate.

Autotaxin is a calcium-dependent ectonucleotide pyrophosphatase and phosphodiesterase. It releases one phosphate group from the substrate ATP to yield PPi, while it simultaneously cleaves the second phosphate group from the substrate ADP by another water molecule to yield AMP. As a result, two molecules of PPi are generated for every single molecule of ATP cleaved by this enzyme.

Autotaxin is predominately regarded as an LPA-producing enzyme that is present in peripheral blood. From here it can be transported to many different types of cells via various secretory pathways. One main function of ATX in production and the LPA signalling is in facilitating the wound healing process. This is further highlighted by the fact that the synthesis and the secretion is elevated in response to inflammatory cytokines and tissue damage.

INTENDED USE

Human autotaxin ELISA kit is intended for analysing the concentration of autotaxin (ATX) present is human samples of serum or plasma.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

1. Microtitre Strips (96 Wells): Coated rabbit polyclonal antibody against human autotaxin.
2. 10x Wash Buffer.
3. 5x Assay Buffer.
4. 100x Detection Antibody Buffer: Labelled biotin polyclonal antibody.
5. Standard: Lyohilized recombinant.
6. 200x STP-HRP Solution.
7. Substrate Solution.
8. Stop Solution.

TYPICAL RESULTS

For the following autotaxin ELISA kit it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 0.78, 1.56, 3.12, 6.25, 12.5, 25, 50 ng/ml.

ASSAY CHARACTERISTICS

– Sensitivity: 0.78 ng/ml.
– Assay Range: 0.78 – 50 ng/ml.
– Specificity: No interference or cross reaction observed.
– Inter-Assay Precision: 5% (CV).
– Intra-Assay Precision: 6.1% (CV).
– Linearity: 100% (1:40 dilution), 102.8-105.6% (1:80 dilution).

REFERENCES

1. Autotaxin: An Early Warning Biomarker for Acute-on-chronic Liver Failure. J Clin Transl Hepatol. (2020) 8 (3): 240-245. Nie C., et al.
2. The autotaxin-lysophosphatidic acid pathway mediates mesenchymal cell recruitment and fibrotic contraction in lung transplant fibrosis. J Heart Lung Transplant. (2021) 40 (1): 12-23. Sinclair K.A., et al.
3. Serum ATX as a novel biomarker for breast cancer. Medicine (Baltimore). (2019) 98 (13): e14973. Shao Y., et al.
4. Evaluation of serum ATX and LPA as potential diagnostic biomarkers in patients with pancreatic cancer. BMC Gastroenterol. (2021) 21 (1): 58. Chen J., et al.

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