1. What was the purpose of washing the red blood cells with saline prior to lysis?
2. What property of the lysis buffer caused the red blood cells to lyse?
3. What are the differences between the primary and secondary antibodies usedin this study?
4. What are the advantages of using indirect immunodetection over directimmunodetection?
5. Antibodies are made when animals detect a foreign antigen. All mammalscarry albumin in their blood. How was a rabbit antibody against albumincreated? Was this lethal to the rabbit? Explain you reasoning.
6. Albumin is not a cytoskeletal protein. What was the purpose of usingantibodies to detect albumin in our cytoskeletal preparations?
7. Why was it necessary to transfer the proteins separated by SDS-PAGE ontonitrocellulose membranes prior to incubation with anti-albumin antibodies?
8. Why did we electrophoretically transfer the proteins resolved by SDS-PAGEonto nitrocellulose? Can you suggest another way to transfer proteins from aSDS gel to a membrane?
9. Why did we wash the nitrocellulose blots with blotting buffer three times afterincubating with the primary and secondary antibodies?