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How do Elisa Kit Work?

Enzyme-linked immunosorbent assay (ELISA) is a widely used analytical method for detecting and quantifying antigens (proteins, hormones, toxins, etc.) in biological fluids, such as serum or plasma. The basic principle of ELISA involves the use of antibodies that specifically recognize and bind to a target antigen.
Here's how ELISA works:
1.Coating: The first step is to coat the bottom of a well in a microtiter plate with a specific antigen of interest. This well acts as a "trap" for the antigen-antibody reaction
2.Blocking: Next, the plate is blocked with a solution to prevent nonspecific binding of proteins to the well. This helps ensure that only the target antigen will bind to the well.
3.Adding the sample: The biological fluid containing the target antigen is added to the well. If the target antigen is present, it will bind to the antigen coating the well.
4.Adding the primary antibody: A primary antibody specific for the target antigen is then added to the well. The antibody is linked to an enzyme that can produce a detectable signal, such as a color change. The primary antibody will bind to the target antigen if it is present in the sample.
5.Detecting the signal: After washing the well to remove any unbound antibodies, a substrate for the enzyme is added. If the target antigen is present in the sample, it will bind to the primary antibody, which in turn will generate a signal. The signal is usually measured by the intensity of color generated by the substrate-enzyme reaction, which is proportional to the amount of target antigen present in the sample.
6.Interpreting the results: Finally, the optical density of the signal is measured and compared to a standard curve to determine the concentration of the target antigen in the sample.
ELISA is a highly sensitive and specific method for detecting antigens and is widely used in medical diagnosis, research, and other applications.
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