Question

Compare and contrast how antibodies and GFP fusion proteins work to localize proteins.

Answer 1

We cal localize the protein either using antibody or GFP fusion protein.

By using antibody.

Here we used florescent tagged antibodies against our target protein. We will incubate the cell with flouroscencnt labelled antibodies. After incubation, depending on the fluorescent marker used we can localize the protein in the cell by using fluorescent microscopes. By this technique we can only localize the position of a protein we can not track the pathway followed by protein.

In GFP tagged protein, we add a GFP protein at the C terminal part of our protein. Here GFP act as a tag for the target protein. GFP is a fluorescent protein and use of this excludes the requirements of using tagged antibodies in the procedure. As the fused protein synthsized, we can track their movements in to the cellular compartment by looking the GFP flouroscence under the microscope. As well as final location of the protein can also be detected.