Immunoprecipitation of protein from Cell Lysate using Antibody
1. Which form of immunoprecipitation is specifically designed to study protein-DNA interactions?
2. In the preparation of immune complexes, what is the typical ratio of Lysis Buffer to the protein sample?
3. Which type of beads should be used for immunoprecipitation when working with a polyclonal antibody?
4. At what temperature and for how long should the immune complex mixture be incubated?
5. Which buffer is added to the precipitate after centrifuging the Sepharose beads mixture to prepare it for use?
6. What is the purpose of boiling the mixture at 100 degrees Celsius for 5 minutes after adding the Loading Buffer?
7. Which elution buffer would you use if you want to perform a western blot analysis on the isolated proteins?
8. What is the role of the rotary agitator in the immunoprecipitation protocol?
9. Why might the Sepharose beads be washed twice before the buffer is added in the preparation step?
10. Which step ensures the elimination of unbound materials after the incubation period in the immunoprecipitation protocol?