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ILEI (Interleukin-like EMT inducer) /FAM3C

ILEI (Interleukin-like EMT-inducer, also called FAM3C) is a new secreted cytokine-like protein of 25 kDa that has structural homology to four-helix-bundle cytokines such as erythropoietin, growth hormone or interleukine members [1]. There are four genes in this new family : FAM3A, FAM3B, FAM3C (ILEI) and FAM3D [1] . ILEI possess a novel GG domain (because of two well-conserved glycine residues) present in eukaryotic FAM3 family, POMGnT1 (protein O-linked mannose beta-1,2-N-acetylglucosaminyl-transferase), TEM2 proteins as well as phage gp35 proteins [2]. While human FAM3B and D are expressed in a few tissues (such as pancreas and placenta), human FAM3A and FAM3C / ILEI are ubiquitously expressed. Recently, ILEI has been identified as a key player in epithelial-to-mesenchymal transition (EMT), in carcinoma progression as well as in metastasis [3]. ILEI acts downstream of TGFbeta receptor signaling and is regulated at the level of translation during EMT [3]. ILEI expression is increased in many carcinomas and could serve as an independent prognostik marker in breast cancer [3]. ILEI is also secreted by pancreatic cancer cells [4, 5].

ILEI (human) Detection Set (APO-54N-032)

Summary of features

	For the quantitative determination of human ILEI from biological fluids (serum, plasma and cell culture supernatant)
	Monoclonal antibodies-based sandwich ELISA
	Detection limit: 0.2 ng / ml
	Range: 0-20 ng / ml
	Specificity : Detects only human ILEI
	Format: sufficient materials to run ELISAs on 5 x 96-well plates

Principle of the Kit

This assay is a sandwich Enzyme Linked-Immuno-Sorbent Assay (ELISA) developed for the direct measurement of human ILEI (hILEI) in biological fluids. A monoclonal antibody specific for hILEI (COAT) is coated onto the wells of the microtiter plate. Samples and standards of ILEI are pipetted into the wells for binding to the coated antibody. After extensive washing to remove unbound compounds, hILEI is recognized by the addition of a biotinylated monoclonal antibody specific for hILEI (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 hILEI in the samples.

Linearity of the STD curve.

Literature Overview:

1. Zhu, Y., Xu, G., Patel, A., McLaughlin, M.M., Silverman, C., Knecht, K., Sweitzer, S., Li, X., McDonnell, P., Mirabile, R., et al. (2002). Cloning, expression, and initial characterization of a novel cytokine-like gene family. Genomics 80, 144-150.

2. Pilipenko, V.V., Reece, A., Choo, D.I., and Greinwald, J.H., Jr. (2004). Genomic organization and expression analysis of the murine Fam3c gene. Gene 335, 159-168.

3. Waerner, T., Alacakaptan, M., Tamir, I., Oberauer, R., Gal, A., Brabletz, T., Schreiber, M., Jechlinger, M., and Beug, H. (2006). ILEI: a cytokine essential for EMT, tumor formation, and late events in metastasis in epithelial cells. Cancer cell 10, 227-239.

4. Gronborg, M., Kristiansen, T.Z., Iwahori, A., Chang, R., Reddy, R., Sato, N., Molina, H., Jensen, O.N., Hruban, R.H., Goggins, M.G., et al. (2006). Biomarker discovery from pancreatic cancer secretome using a differential proteomic approach. Mol Cell Proteomics 5, 157-171.

5. Mauri, P., Scarpa, A., Nascimbeni, A.C., Benazzi, L., Parmagnani, E., Mafficini, A., Della Peruta, M., Bassi, C., Miyazaki, K., and Sorio, C. (2005). Identification of proteins released by pancreatic cancer cells by multidimensional protein identification technology: a strategy for identification of novel cancer markers. Faseb J 19, 1125-1127.