What is an Enzyme-Linked Immunosorbent Assay (ELISA)?
The ELISA is a method to determine the concentrations (on the order of ng/mL to pg/mL) of a material in a solution. The ELISA will measure optical density and using that information, you can determine the concentration of the protein of interest. In order to do this, you can create a standard curve graph for the optical density versus the concentration of the protein; extrapolation from this curve will yield the concentration of the protein. For more information on this tool, click here. How is HRP used in an ELISA? HRPs are used as secondary antibodies in ELISAs. Therefore, the HRP will bind to a primary antibody, which binds to the target protein (the target protein is the protein that you wish to detect). The HRP will then bind to a substrate to yield a signal that leads to detection; HRP catalyzes a reaction with the substrate that causes the color change (Schutz and others 1997). Figure 2. How HRP might work in an ELISA (or Western Blot). In this example, the primary antibody has
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- What is an Enzyme-Linked Immunosorbent Assay (ELISA)?
