Name the experiment shown above and briefly describe how it is set up as well as the role of each component in that setup. What is it used for?
Gel electrophoresis is a method of separation of protein (SDS-PAGE, polyacrylamide gel electrophoresis) and DNA (agarose gel electrophoresis) molecule. In the given question, gel picture is for the protein sample and its interaction is shown with a labeled probe and antibody.
SDS-PAGE is used to separate the protein molecules based on its sizes when it migrates towards the cathode. All proteins will be linearized and negatively charged with SDS molecules. Smaller protein will move faster and be observed in the bottom of the gel while larger protein will be present in the top of the gel as it moves slowly.
Western blotting is a method to transfer the proteins on the nitrocellulose membrane and observe the protein using primary and secondary antibodies against that protein. Through this method, we can check very small quantities of proteins which we cannot observe by the normal SDS-PAGE staining method. Secondary antibody will be labeled with the enzyme and substrate addition will give color change
Electrophoretic mobility shift assay - EMSA is a method to study the interaction of between DNA and proteins. Because we know DNA is a genetic material and it interacts with enzymes (proteins) while replication, recombination, transcription and translation. So, through this method we can resolve complexes of DNA-protein with different stoichiometry or conformation. We can use both crude protein samples or purified one, both will work.
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