proteins (antigens) are separated by size or electrophoresis gel and detected by antibody "sandwich" using radioactivity or visible light detection
(ex: used to confirm a positive HIV antibody ELISA test); present antibodies will stick... color response. End result:
bands. Does the patient respond to one, two or three proteins from microbe. ex: HIV has three proteins..
The basic technique of a Western blot involves sorting proteins by length on a gel and then probing the gel with antibodies that react to the proteins that are being searched for. However, when Western blots are used for HIV testing, the process is actually performed in reverse. Instead of unknown proteins being tested for with known antibodies, labs work with prepared protein samples, and look to see if there are any antibodies in a person's blood that stick to them.)
- 1.Proteins from a known bacterium or virus are separated by an electric current in electrophoresis
- 2. The proteins are then transferred to a filter by blotting.
- 3. Patients serum is washed over the filter. IF the patient has antibodies to one of the proteins in the filter then antiboidies and protein will combine. Anti human serum linked to an enzyme is then washed over the filter.
- 4. THis will be made visible as a colored band on the filter after addition of the enzyme's substrate