You are provided with a sample of blood from a patient who is suspected of having Lymes disease (Borrelia). If you also have access to Borrelia cell surface proteins and goat anti-human IgG conjugated horse radish peroxidase state how you would determine if the individual had been exposed to this disease.
This is a great question to test your technical knowledge of Western Blotting. Just to review quickly, Western Blots are a method of detecting one specific protein of interest out of a mixture of proteins. The steps involved in Western Blotting are as follows:
1. Separate the mixture of proteins using SDS-PAGE (polyacrylamide gel electrophoresis).
2. Transfer the proteins from the gel to a nitrocellulose membrane using another electrophoresis.
3. Incubate the nitrocellulose membrane with a primary antibody that will specifically bind and recognize the protein you are interested in.
4. Incubate the nitrocellulose membrane with a secondary antibody that is conjugated (linked) to an enzyme. This antibody will recognize and bind to the primary antibody-protein of interest complex. The enzyme that is bound to the secondary antibody ...
This solution reviews in 542 words the technique of Western Blotting, including a definition and step-by-step breakdown of the technique in general as well as in the application for detecting Lyme's disease.