DcR3 Human ELISA (soluble Decoy Receptor 3, TR6, M68)

Research topic

Oncology

Features

• The total assay time is less than four hours.
• The kit measures total serum Decoy Receptor 3.
• Calibrator is purified native protein based.
• Quality Controls are human serum based.
• Components of the kit are in the lyophilized, concentrated and ready-to-use states.

Storage/Shipping

Store the kit at 2–8°C. Under these conditions, the kit is stable until the expiration date (see label on the box).

Summary

The Human Decoy Receptor 3 ELISA is a highly specific double monoclonal sandwich enzyme immunoassay for the quantitative measurement of human soluble Decoy Receptor 3 in serum, plasma and tissue culture medium. A decoy receptor, DcR3 (also known as TR6 and M68), belongs to the tumor necrosis factor receptor (TNFR) superfamily . DcR3 is a 33 kDa glycoprotein consisting of 300 amino acids and shares sequence identity with OPG (31%), TNFRII (29%) and Fas (17%). DcR3 binds to at least three different ligands, Fas ligand (FasL), LIGHT, and TL1A, and the biological functions these three ligands are blocked by the presence of DcR3. DcR3 might act as a modulator in the immune system. A soluble recombinant DcR3 was shown to modulate T-cell responses as well as differentiation and maturation of dendritic cells and, subsequently, the TH2 response. DcR3 was found in lung tissue, tumor cells and in endothelial cells. DcR3 expression was localized in human lung carcinomas, gastrointestinal tract tumors, virusassociated lymphomas (Epstein-Barr virus or HTLV-1), malignant gliomas and glioma cells, malignant melanoma, silicosis, and systemic lupus erythematosus. DcR3 also may play a role in the pathogenesis of allergic diseases.

Assay format

Sandwich ELISA, Biotin-labelled antibody

Sample requirements

35 µl/well

Applications

Cell culture medium, Plasma-Citrate, Plasma-EDTA, Plasma-Heparin, Serum

Calibration Curve

Calibration range

0.5 – 12 ng/ml

Limit of detection

Analytical Limit of Detection is calculated from the real Decoy Receptor 3 values in wells and is 0.033 ng/ml

Limit of quantification

Assay Sensitivity takes the dilution of samples into consideration and is calculated according to the formula: Assay Sensitivity = Analytical Limit of Detection x sample dilution = 0.033 ng/ml x 3 = 0.1 ng/ml

Intra-assay (Within-Run, n=8)

CV = 3.5 %

Inter-assay (Run-to-Run, n=5)

CV = 6.5 %

Spiking Recovery

88.3 %

Dilution Linearity

105.7 %

Cross-Reactivity

Human

References to summary

• Wu Y., Han B., Sheng H., Lin M., Moore P. A., Zhang J., Wu J.: Clinical significance of detecting elevated serum DcR3/TR6/M68 in malignant tumor patients. Int. J Cancer, 105 (5): 724–732 (2003)
• Chen Ch. Ch., Yang Y. S., Lin Y. T., Hsieh S. L., Chiang B-L.: Soluble decoy receptor 3: Increased levels in atopic patients. J. Allergy Clin Immunol, 114(1): 195–197 (2004)
• Wu Y. Y., Chang Y. C., Hsu T. L., Hsieh S. L., Lai M. Z.: Sensitization of cells to TRAIL-induced apoptosis by decoy receptor 3. J. Biol Chem, 279 (42): 44211–44218 (2004)
• Pitti R. M., Marsters S. A., Lawrence D. A., Roy M., Kischkel F. C., Dowd P., Huang A., Donahue C. J., Sherwood S. W., Baldwin D. T., et al.: Genomic amplification OF a decoy receptor for Fas ligand in lung and colon cancer. Nature, 396: 699–703 (1998)
• Hsu T. L., Chang Y. C., Chen S. J., Liu Y. J., Chiu A. W., Chio C. C., Chen L., Hsieh S. L.: Modulation of dendritic cell differentiation and maturation by decoy receptor 3. J Immunol, 168: 4846–4853 (2002)
• Bai C., Connoly B., Metzker M. L., Hilliard C. A.., Liu X., Sanding V., Soderman A., Galloway S. M., Liu Q., Austin C. P., et al.: Overexpression OF M68/DcR3 in human gastrointestinal tract tumors independent OF gene amplification and its location in a four-gene cluster. Proc. Natl. Acad. Sci. USA, 93 (3): 1230–1235 (2000)
• Yu, K. Y., Kwon B., Ni J., Zhai Y., Ebner R., Kwon B.S.: A newly identified member of tumor factor receptor superfamily (TR6) suppresses LIGHT-mediated apoptois. J Biol. Chem., 274: 13733–13736 (1999).
• Sung H. H., Juang J. H., Lin Y. C., Kuo C. H., Hung J. T., Chen A., Chang D. M., Chang S. Y., Hsieh S. L., Sytwu H. K.: Transgenic expression of decoy receptor 3 protect islets from spontaneous and chemical-induced autoimmune destruction in nonobese diabetic mice. J. Exp Med, 199 (8): 1143–1151 (2004)