Autotaxin ELISA Kit for Mouse

Background: Autotaxin, also known as ENPP-2, is a secreted glycoprotein which belongs to the ectonucleotide pyrophosphatase/phosphodiesterase (NPP) family [1,2]. Generally, NPPs can hydrolyze phosphates from nucleotides. Autotaxin exhibits the unique lysophospholipase D activity [3]. The mature protein includes two somatomedin-B-like (SMB) cysteine knot domains, a catalytic domain, and an inactive C-terminal nuclease-like domain with an EF-handlike motif that is important in cell motility, and a region involved in autotaxin secretion [1,4,5]. There are three isoforms identified in mouse and human [6,7]. Most circulating autotaxin is the beta form which contains 863 amino acid. Autotaxin contributes to the predominant extracellular source of the phospholipid LPA (lysophosphatidic acid) from LPC (lysophosphatidylcholine) [8-11]. Autotaxin can also produce minor amounts of sphingosine 1-phosphate and cyclic phosphatidic acid which can antagonize many of the tumorigenic properties of LPA [9,12]. Autotaxin stimulates tumor cell motility and enhances invasion and metastasis. It's upregulated in melanoma, glioblastoma, breast and lung carcinoma, follicular lymphoma and other cancers [2,8,11]. Autotaxin production by adipocytes enhances pre-adipocyte proliferation and may be elevated in obesity [11, 13]. Autotaxin is present in blood, urine, saliva, seminal and cerebrospinal fluids [2, 3]. In addition, plasma autotaxin is cleared by the liver which is elevated in liver disease [3, 14]. Normal serum or plasma autotaxin concentration is reported to be slightly higher in females than in males, and highest in pregnant females [2, 14].

Principle of the Assay: This assay is a quantitative sandwich ELISA.The micro-plate is pre-coated with a rabbit polyclonal antibody against mouse autotaxin. Standards and samples are pipetted into the wells and any mouse autotaxin present is bound by the immobilized antibody. After washing away any unbound substances, a biotin labelled polyclonal antibody specific for mouse autotaxin is added to the wells. After wash step to remove any unbound reagents, streptavidin-HRP conjugate (STP-HRP) is added. After the last wash step, an HRP substrate solution is added and colour develops in proportion to the amount of mouse autotaxin bound initially. The assay is stopped and the optical density of the wells determined using a microplate reader. Since the increases in absorbance are directly proportional to the amount of captured mouse autotaxin, the unknown sample concentration can be interpolated from a reference curve included in each assay

Intended Uses: This mouse autotaxin ELISA kit is designed for quantification of mouse autotaxin in serum, plasma and cell culture supernate samples.

Cimpean, Anisoara, et al. "Substrate-specifying determinants of the nucleotide pyrophosphatases/phosphodiesterases NPP1 and NPP2." Biochemical Journal1 (2004): 71-77.
Okudaira, Shinichi, Hiroshi Yukiura, and Junken Aoki. "Biological roles of lysophosphatidic acid signaling through its production by autotaxin." Biochimie6 (2010): 698-706.
Umezu-Goto, Makiko, et al. "Autotaxin has lysophospholipase D activity leading to tumor cell growth and motility by lysophosphatidic acid production." J cell biol2 (2002): 227-233.
Hausmann, Jens, et al. "Structural basis of substrate discrimination and integrin binding by autotaxin." Nature Structural and Molecular Biology 18.2 (2011): 198.
Dennis, Jameel, et al. "Phosphodiesterase-Ialpha/autotaxin's MORFO domain regulates oligodendroglial process network formation and focal adhesion " Molecular and Cellular Neuroscience 37.2 (2008): 412-424.
van Meeteren, Laurens A., and Wouter H. Moolenaar. "Regulation and biological activities of the autotaxin-LPA axis." Progress in lipid research2 (2007): 145-160.
Giganti, Adeline, et al. "Murine and Human Autotaxin alpha, beta, and gamma Isoforms GENE ORGANIZATION, TISSUE DISTRIBUTION, AND BIOCHEMICAL CHARACTERIZATION." Journal of Biological Chemistry12 (2008): 7776-7789.
Gijsbers, Rik, et al. "The hydrolysis of lysophospholipids and nucleotides by autotaxin (NPP2) involves a single catalytic site." FEBS letters1-3 (2003): 60-64.
Tsuda, Satomi, et al. "Cyclic phosphatidic acid is produced by autotaxin in blood." Journal of Biological Chemistry 281.36 (2006): 26081-26088

Small volumes of Autotaxin elisa kit vial(s) may occasionally become entrapped in the seal of the product vial during shipment and storage. If necessary, briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container`s cap. Certain products may require to ship with dry ice and additional dry ice fee may apply.

MBS398111 is a ready-to-use microwell, strip-or-full plate ELISA (enzyme-linked immunosorbent assay) Kit for analyzing the presence of the Autotaxin, ELISA Kit target analytes in biological samples. The concentration gradients of the kit standards or positive controls render a theoretical kit detection range in biological research samples containing Autotaxin. The ELISA analytical biochemical technique of the MBS398111 kit is based on Autotaxin antibody-Autotaxin antigen interactions (immunosorbency) and an HRP colorimetric detection system to detect Autotaxin antigen targets in samples. The ELISA Kit is designed to detect native, not recombinant, Autotaxin. Appropriate sample types may include undiluted body fluids and/or tissue homogenates, secretions. Quality control assays assessing reproducibility identified the intra-assay CV (%) and inter-assay CV(%).