AAT Bioquest

What are the detection methods for western blot?

Posted October 25, 2022


The main detection methods for western blot are the following: fluorescence detection, chemiluminescent detection, colorimetric detection. Chemiluminescence detection occurs when a substrate is catalyzed by enzymes and produces light as a byproduct. The limiting reagent is a substrate, and light production stops when the substrate is used up. Fluorimetric detection requires the use of an antibody that has been labeled with a fluorophore. The fluorophore is then excited with a light and becomes emitted. The emitted light produces a higher wavelength for detection. Colorimetric detection is dependent on the creation of a colored substance that moves onto the western blot. It is formed after the conversion of a chromogenic blotting substrate by an enzyme. These experiments are ideal for the detection of many proteins since the reaction does not require a machine to read and is highly stable.

Additional resources

Western Blotting Assays

HRP-labeled goat anti-mouse IgG (H+L)

Colorimetric detection