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Experimental analysis of zebrafish triiodothyronine (T3)
The Zebrafish Triiodothyronine (T3) Experimental Analysis aims to quantify the levels of triiodothyronine (T3) in zebrafish serum, plasma, and other related biological fluids. This assay is based on the double-antibody sandwich immunoassay method, which ensures high specificity and accuracy. The process begins by coating a microplate with purified zebrafish T3 antibodies, creating a solid-phase antibody matrix. After incubation, the sample containing T3 antigen is added, allowing it to bind to the immobilized antibodies. Subsequently, an HRP-conjugated anti-T3 antibody is introduced, forming a complex of antibody-antigen-enzyme-labeled antibody. Following thorough washing to remove unbound components, a TMB substrate is added. Under the catalytic action of HRP, TMB turns blue, and upon acid termination, it changes to a yellow color. The intensity of the color is directly proportional to the concentration of T3 in the sample. The optical density (OD) at 450 nm is measured using a microplate reader, and the T3 concentration is determined by comparing the OD value to a pre-established standard curve. This method provides a reliable and efficient way to assess thyroid hormone levels in zebrafish, supporting research in developmental biology, endocrinology, and toxicology.