Thyroxine Experimental Analysis Description
Purpose: This kit is designed to measure the concentration of thyroxine in zebrafish serum, plasma, and other related liquid samples. It provides a reliable and accurate method for assessing thyroid hormone levels, which is essential for studying endocrine function and environmental toxicology in zebrafish models.
Experimental Principle: The assay is based on the double-antibody sandwich immunoassay technique. A microtiter plate is pre-coated with purified zebrafish thyroxine-specific antibodies, forming a solid-phase antibody. After adding the sample, thyroxine present in the sample binds to the immobilized antibody. Subsequently, an HRP-conjugated thyroxine antibody is added, forming a complex of antibody-antigen-enzyme-labeled antibody. Following thorough washing to remove unbound components, a TMB substrate is introduced. Under the catalytic action of HRP, TMB turns blue and then changes to yellow when an acidic stop solution is added. The intensity of the color is directly proportional to the thyroxine concentration in the sample.
The absorbance is measured at 450 nm using a microplate reader, and the thyroxine concentration is determined by comparing the sample's OD value to a standard curve generated from known concentrations. This method ensures high specificity and sensitivity, making it ideal for quantitative analysis of thyroxine in zebrafish biological samples.
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