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.© BrainMass Inc. brainmass.com October 24, 2018, 5:29 pm ad1c9bdddf
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.
Microbiology Study Questions
21. Explain the purpose of Bergey's Manual and differentiate the types.
22. Describe how staining and biochemical tests are used to identify bacteria.
23. Differentiate between Western blotting and Southern blotting.
24. Explain how serological tests and phage typing can be used to identify an unknown organism.
25. Describe how a new bacterium can be classified by the following molecular methods: DNA base composition, rRNA sequencing, DNA fingerprinting, PCR, and nucleic acid hybridization.
26. Describe the defining characteristics of cyanobacteria, fungi, platyhelminths, cestodes, houseflies, algae, archaezoa, microspora, yeasts, helminths, protozoa
27. Differentiate between an intermediate host and a definitive host and give an example of what parasite stage is seen in humans as intermediate and definitive hosts.
28. Describe the defining characteristics of Bacteroides, Thiobacillus, Bacillales, Neisseria, Pseudomonas.
29. Differentiate between a tick and a mosquito, and name a disease transmitted by each.
30. Discuss the relationship of DNA- and RNA-containing viruses to cancer.