The experimental principle of this kit is based on the double-antibody sandwich ELISA method, which is used to quantify human myostatin (MSTN) in biological samples. The process begins with a microplate that has been pre-coated with a specific monoclonal antibody against human MSTN. This coated antibody acts as a solid-phase capture agent. After adding the sample, any MSTN present will bind to the immobilized antibody. Next, a horseradish peroxidase (HRP)-labeled secondary antibody, which recognizes a different epitope on MSTN, is introduced. This forms a complex consisting of the captured MSTN, the primary antibody, and the HRP-conjugated secondary antibody. Following a thorough wash to remove unbound components, a substrate solution containing TMB (3,3',5,5'-tetramethylbenzidine) is added. Under the catalytic action of HRP, TMB is oxidized and turns blue. When an acidic stop solution is added, the color changes from blue to yellow. The intensity of the yellow color is directly proportional to the concentration of MSTN in the sample. The optical density (OD) at 450 nm is then measured using a microplate reader. By comparing the OD values of the test samples with those of a standard curve generated from known concentrations of MSTN, the exact level of MSTN in the sample can be accurately determined. This method ensures high specificity, sensitivity, and reproducibility for MSTN detection.
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