| Amyloid P component, serum (SAP), also known as PTX2 or APCS, is the identical serum form of amyloid P component (AP), a 25kD pentameric protein first identified as the pentagonal constituent of in vivo pathological deposits called amyloid. It belongs to the pentraxins family, characterised by calcium dependent ligand binding and distinctive flattened beta-jellyroll structure similar to that of the legume lectins. This gene is mapped to 1q23.2. The binding of the encoded protein to proteins in the pathological amyloid cross-beta fold suggests its possible role as a chaperone. This protein is also thought to control the degradation of chromatin. It has been demonstrated that this protein binds to apoptotic cells at an early stage, which raises the possibility that it is involved in dealing with apoptotic cells in vivo. The Amyloid P Component, Serum (SAP/PTX2) BioAssay(TM) ELISA Kit is for quantitative detection of mouse SAP/PTX2 in cell lysates, cell culture supernates, serum, plasma (Heparin or EDTA). Detection Range: 1.56-100ng/ml Sensitivity: <50pg/ml Specificity: Recognizes natural and recombinant mouse SAP/PTX2. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This Amyloid P Component, Serum (SAP/PTX2) BioAssay(TM) ELISA Kit is a Sandwich High Sensitivity ELISA is a solid phase immunoassay specially designed to measure mouse SAP/PTX2 with a 96-well strip plate that is pre-coated with antibody specific for SAP/PTX2. The detection antibody is a biotinylated antibody specific for SAP/PTX2. The capture antibody is a monoclonal from rat and the detection antibody is a polyclonal from goat. The kit contains recombinant mouse SAP/PTX2 with immunogen: Expression system for standard: NS0, Immunogen sequence: Q21-D224. To measure mouse SAP/PTX2, 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 density of the yellow product is linearly proportional to mouse SAP/PTX2 in the sample. Read the density 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 mouse SAP/PTX2 in the sample. Kit Components: 353028A: Microtiter Strips, 1x96 wells *353028B: SAP/PTX2 Standard, 2x100ng 353028C: Anti-mouse SAP/PTX2 (Biotin), 1x130ul (1:100 dilution) 143696: Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (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 *353028B 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. |
