| The ST2, Mouse BioAssay(TM) ELISA Kit is a 4.5 hour solid phase ELISA designed to measure mouse ST2 in cell culture supernates, serum, and plasma. Test Principle: This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for mouse ST2 has been pre-coated onto a microplate. Standards, Control, and samples are pipetted into the wells and any mouse ST2 present is bound by the immobilized antibody. After washing away any unbound substances, an enzyme-linked monoclonal antibody specific for mouse ST2 is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells. The enzyme reaction yields a blue product that turns yellow when the Stop Solution is added. The intensity of the color measured is in proportion to the amount of mouse ST2 bound in the initial step. The sample values are then read off the standard curve. Kit Components: S6606-50P1: Microwell Strips, 1x96 wells *S6606-50P2: Standard (lyophilized), 1x1vial (50ng) *S6606-50P3: Control (lyophilized), 0.97-1.62ng/ml, 1x1vial S6606-50P4: HRP Conjugate, 1x12ml S6606-50P5: Assay Diluent, 1x12ml S6606-50P6: Calibrator Diluent, 1x21ml S6606-50P7: Wash Buffer, 25X, 1x50ml S6606-50P8: Color Reagent A, 1x12ml S6606-50P9: Color Reagent B, 1x12ml S6606-50P10: Stop Solution (HCl), 1x23ml Storage and Stability: Store components at 4C. Stable for 6 months after receipt. Upon reconstitution of *S6606-50P2 & *S6606-50P3, aliquot and store at -20C for up to 1 month. Avoid repeated freeze-thaw cycles. For maximum recovery of product, centrifuge the original vial prior to removing the cap. Sensitivity: Fifty-three assays were evaluated and the minimum detectable dose (MDD) of mouse ST2 ranged from 0.004-0.059ng/ml. The mean MDD was 0.014ng/ml. The MDD was determined by adding two standard deviations to the mean optical density value of twenty zero standard replicates and calculating the corresponding concentration. Specificity: This assay recognizes both recombinant and natural mouse ST2. The factors listed below were prepared at 100ng/ml in Calibrator Diluent (1X) and assayed for crossreactivity. Preparations of the following factors prepared at 100ng/ml in a mid-range mouse ST2 control were assayed for interference. No significant crossreactivity or interference was observed. Recombinant mouse: IL-1a, IL-1b, IL-1ra, IL-1 R1, IL-1 R2, IL-1 RAPL2/IL-1 R9, IL-1 Rrp2/IL-1 R6, IL-18, IL-33 (aa1-108), SIGIRR, TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR12 Recombinant mouse IL-33 (aa109-266) does not crossreact in this assay, but it does interfere at concentrations above 0.188ng/ml. Recombinant human: IL-1 RAcp/IL-1 R3, IL-33 (aa1-111), IL-33 (aa112-270), ST2/IL-1 R4, TIGIRR |