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The final step of Western blotting is detection; after the enzyme-linked secondary antibody has bound, adding the corresponding substrate causes a change in color or fluorescence that signals the presence of the target protein.
After preparing the chromogenic or chemiluminescent substrate according to the instructions on the datasheet, cover the membrane with the substrate and incubate on a shaker until bands appear. Rinse the membrane and cover with film or a glass cover. Photograph the gel or expose the film and enjoy the excellent results of using Boster products.
For a detailed protocol for signal development, see our Western blot handbook.
Chemiluminescent development tends to be more sensitive, and more expensive than chromogenic development. Be sure to pick the right method for your needs.
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|Applications||To detect antigen at low picogram level by reacting with horseradish peroxidase|