| Death receptor 3 (DR3), also known as tumor necrosis factor receptor superfamily member 25 (TNFRSF25), is a cell surface receptor of the tumor necrosis factor receptor superfamily which mediates apoptotic signalling and differentiation. Knockout studies in mice suggested the role of this gene in the removal of self-reactive T cells in the thymus. Multiple alternatively spliced transcript variants of this gene encoding distinct isoforms have been reported, most of which are potentially secreted molecules. The alternative splicing of this gene in B and T cells encounters a programmed change upon T-cell activation, which predominantly produces full-length, membrane bound isoforms, and is thought to be involved in controlling lymphocyte proliferation induced by T-cell activation. The TNFRSF25/DR3 BioAssay(TM) ELISA Kit is a Sandwich High Sensitivity BioAssay(TM) ELISA kit quantitative detection of human TNFRSF25/DR3 in cell lysates, cell culture supernates, serum, plasma (Heparin or EDTA). Detection Range: 312pg/ml-20,000pg/ml Sensitivity: <10pg/ml Specificity: Recognizes natural and recombinant Human TNFRSF25/DR3. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This TNFRSF25/DR3 BioAssay(TM) ELISA Kit is a Sandwich High Sensitivity ELISA is a solid phase immunoassay specially designed to measure human TNFRSF25/DR3 with a 96-well strip plate that is pre-coated with antibody specific for TNFRSF25/DR3. The detection antibody is a biotinylated antibody specific for TNFRSF25/DR3. The kit contains recombinant human TNFRSF25/DR3 with immunogen: Expression system for standard: NS0, Immunogen sequence: Q25-F201. To measure human TNFRSF25/DR3, add standards and samples to the wells, then add the biotinylated detection antibody. Wash the wells with PBS buffer, and add Avidin-Biotin-Peroxidase Complex (ABC-HRP). Wash away the unbounded ABC-HRP with PBS buffer and add TMB. TMB is substrate to HRP and will be catalyzed to produce a blue color product, which changes into yellow after adding acidic stop solution. The absorbance of the yellow product at 450nm is linearly proportional to human TNFRSF25/DR3 in the sample. Read the absorbancey of the yellow product in each well using a plate reader, and benchmark the sample wells readings against the standard curve to determine the concentration of human TNFRSF25/DR3 in the sample. Kit Components: 396799A: Microtiter Strips, 1x96 wells *396799B: TNFRSF25/DR3 Standard, 2x20ng 396799C: Anti-human TNFRSF25/DR3 (Biotin), 1x100ul (1:100 dilution) 143696: Avidin-Biotin-Peroxidase Complex (ABC), 1x100ul (1:100 dilution) 143697: Sample Diluent Buffer, 1x30ml 143698: Antibody Diluent Buffer, 1x12ml 143699: ABC Diluent Buffer, 1x12ml 143700: TMB Color Developing Agent, 1x10ml 143701: TMB Stop Solution, 2N H2SO4, 1x10ml 143702: Wash Buffer, 25X, 1x20ml Storage and Stability: Store *396799B powder at 4C. Once reconstituted store at 4C for up to 12 hours or at -20C for up to 48 hours. Store other components at 4C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Assay Summary: 1. Add 100ul of samples and standards and incubate the plate at 37C for 90 minutes or at RT for 2 hours. Do not wash. 2. Add 100ul biotinylated antibody and incubate the plate at 37C for 60 minutes or at RT for 90 minutes. Wash plate 3 times with Working Wash Buffer. 3. Add 100ul of ABC working solution and incubate the plate at 37C for 30 minutes or at RT for 40 minutes. Wash plate 5 times with Working Wash Buffer. 4. Add 90ul of TMB color developing agent and incubate the plate at 37C in dark for 15-25 minutes or at RT for 30 minutes. 5. Add 100ul TMB Stop Solution and read. |
