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Netrin-4

The Netrins (whose name means “one who guides” in Sanskrit) constitutes a family of laminin-related secreted protein. In mice, human and rat, Netrin-1, Netrin-3/NTL2, Netrin-4/beta and G-Netrins have been cloned [1]. The Netrins are involved in axonal guidance, are required for regeneration and maintenance of the central nervous system and have been implicated in the development of mammary gland, lung, pancreas, and blood vessel [1, 2]. Netrin-4 (also called beta-Netrin) is a novel Netrin more closely related to laminin beta chain that is broadly expressed in both neural and non-neural tissues [3]. It acts during the development and controls the migration of neurons [4]. The role of Netrin-4 in guiding neurones seems to be mediated by known Netrin-1 receptors : DCC and UNC5H1 [5]. Netrin-4 gene (NTN4) is transcribed in a variety of cells including epithelial cells and vascular endothelial cells [4]. Recent studies suggest a role of Netrin-1 and 4 in the branching morphogenesis in the lung [2]. Netrin-4 promotes angiogenesis by stimulating proliferation and migration of microvascular endothelial cells to the same extent that vascular endothelial growth factor (VEGF) [6]. Kidney also express Netrin-4 whose level is down regulated upon ischemia-reperfusion injury [7]. Netrin-4 has been reported to be a novel prognostic marker in breast cancer [8].

Netrin-4 (human) ELISA Kit (APO-54N-037)

Summary of features

	For the quantitative determination of human Netrin-4 from biological fluids (serum, plasma and cell culture supernatant)
	Monoclonal antibodies-based sandwich ELISA
	Detection limit: 50 pg / ml
	Range: 0-5 ng / ml
	Specificity : Detects only human Netrin-4
	Format: 96 wells (~80 tests)

Principle of the Kit

This assay is a sandwich Enzyme Linked-Immuno-Sorbent Assay (ELISA) developed for the direct measurement of human Netrin-4 (hNetrin-4) in biological fluids. A monoclonal antibody specific for hNetrin-4 (COAT) is coated onto the wells of the microtiter plate. Samples and standards of hNetrin-4 are pipetted into the wells for binding to the coated antibody. After extensive washing to remove unbound compounds, hNetrin-4 is recognized by the addition of a biotinylated monoclonal antibody specific for hNetrin-4 (DET). After removal of excess biotinylated antibody, streptavidine-peroxidase is added. Following a final washing, peroxidase activity is quantified using the substrate 3,3’,5,5’-tetramethylbenzidine (TMB). The intensity of the color reaction is measured at 450 nm after acidification and is directly proportional to the concentration of hNetrin-4 in the samples.

Linearity of the STD curve.

Literature Overview:

1. Cirulli, V., and Yebra, M. (2007). Netrins: beyond the brain. Nature reviews 8, 296-306.

2. Liu, Y., Stein, E., Oliver, T., Li, Y., Brunken, W.J., Koch, M., Tessier-Lavigne, M., and Hogan, B.L. (2004). Novel role for Netrins in regulating epithelial behavior during lung branching morphogenesis. Curr. Biol. 14, 897-905.

3. Barallobre, M.J., Pascual, M., Del Rio, J.A., and Soriano, E. (2005). The Netrin family of guidance factors: emphasis on Netrin-1 signalling. Brain Res. Rev. 49, 22-47.

4. Koch, M., Murrell, J.R., Hunter, D.D., Olson, P.F., Jin, W., Keene, D.R., Brunken, W.J., and Burgeson, R.E. (2000). A novel member of the netrin family, beta-netrin, shares homology with the beta chain of laminin: identification, expression, and functional characterization. J. Cell Biol. 151, 221-234.

5. Qin, S., Yu, L., Gao, Y., Zhou, R., and Zhang, C. (2007). Characterization of the receptors for axon guidance factor netrin-4 and identification of the binding domains. Mol. Cell. Neur. 34, 243-250.

6. Wilson, B.D., Ii, M., Park, K.W., Suli, A., Sorensen, L.K., Larrieu-Lahargue, F., Urness, L.D., Suh, W., Asai, J., Kock, G.A., et al. (2006). Netrins promote developmental and therapeutic angiogenesis. Science 313, 640-644.

7. Wang, W., Reeves, W.B., and Ramesh, G. (2008). Part 1: Functional Studies Netrin-1 protects against ischemia reperfusion injury of the kidney. Am J Physiol Renal Physiol.

8. Esseghir, S., Kennedy, A., Seedhar, P., Nerurkar, A., Poulsom, R., Reis-Filho, J.S., and Isacke, C.M. (2007). Identification of NTN4, TRA1, and STC2 as prognostic markers in breast cancer in a screen for signal sequence encoding proteins. Clin Cancer Res 13, 3164-3173.