The BioAssay™ ELISA Kit is a 1.5 hour solid-phase Competitive ELISA designed for the quantitative determination of 13,14-dihydro-15-keto-Prostaglandin F2-Alpha (PGFM) (Canine). This kit for research use only, not for therapeutic or diagnostic applications. Kit Detection Range: 0.5-10ng/mL Kit Sensitivity: 0.1ng/mL Assay Principle: 13,14-dihydro-15-keto-Prostaglandin F2-Alpha (PGFM) BioAssay™ ELISA Kit (Canine) utilizes the quantitative Competitive Enzyme Immunoassay technique, utilizing a monoclonal antibody and an conjugate to 13,14-dihydro-15-keto-Prostaglandin F2-Alpha (PGFM) (Canine). The assay sample and buffer are incubated together with the HRP conjugate in a pre-coated plate for one hour. After the incubation period, the wells are decanted and washed five times. The wells are then incubated with a substrate for HRP enzyme. The product of the enzyme-substrate reaction forms a blue colored complex. Finally, a stop solution is added to stop the reaction, which will then turn the solution yellow. The intensity of color is measured spectrophotometrically at 450nm in a microplate reader. The intensity of the color is inversely proportional to the Target Protein concentration since Protein from samples and Protein-HRP conjugate compete for the anti-Protein antibody binding site. Since the number of sites is limited, as more sites are occupied by Protein from the sample, fewer sites are left to bind the Protein-HRP conjugate. A standard curve is plotted relating the intensity of the color (O.D.) to the concentration of standards. The Target Protein concentration in each sample is interpolated from this standard curve. Materials and Equipment Supplied: 1. Microtiter Plate, 96 wells, stripwell 2. Enzyme Conjugate, 6mL, 1 vial 3. Standard A, 0ng/mL, 1 vial 4. Standard B, 0.5ng/mL, 1 vial 5. Standard C, 1.0ng/mL, 1 vial 6. Standard D, 2.5ng/mL, 1 vial 7. Standard E, 5.0ng/mL, 1 vial 8. Standard F, 10ng/mL, 1 vial 9. Substrate A, 6mL, 1 vial 10. Substrate B,
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