This reagent is intended for research use only. The kit is designed to quantitatively determine the concentration of thyroxine (T4) in zebrafish serum, plasma, and related biological samples. The method employed is a double-antibody sandwich immunoassay. In this procedure, a microtiter plate is pre-coated with purified anti-T4 antibodies, forming a solid-phase antibody. After incubation, the sample containing T4 antigen is added, allowing it to bind to the immobilized antibodies. Subsequently, HRP-conjugated anti-T4 antibodies are introduced, forming a complex of antibody-antigen-enzyme-labeled antibody. Following thorough washing to remove unbound components, a TMB substrate solution is added. Under the catalytic action of HRP, TMB turns blue, and upon addition of an acidic stop solution, it changes to yellow. The intensity of the color developed is directly proportional to the T4 concentration in the sample. The optical density (OD) is measured at 450 nm using a microplate reader. A standard curve is then constructed using known concentrations of T4, enabling the accurate calculation of T4 levels in the test samples. This assay provides a sensitive, specific, and reliable method for T4 detection in zebrafish research.
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