BIOL 300 Lecture Notes - Protein Purification, Secretion, Elution
15 Dec 2012
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Document Summary
Biochemical analysis of signal transduction pathways were perhaps the first used experimentally. It was discovered that in addition to the minimal medium (the bare necessities of all cells, like amino acids, sugars, etc. ), you also needed growth factors to to grow cells in culture. We can see here a ligand (epo) in red binding to a receptor which is a heterodimer (blue and green). You then need to have an activity test, which would be testing the sample extract on a tissue culture to see how well they let the tissue culture cells grow. In reality, people often carry out both steps, first running through an ion-exchange chromatography to get only proteins of a certain charge, and then running that elution through a gel-filtration chromatography to further separate based on size. You can then run the final run on an sds gel to see the exact sizes of the proteins: proteins are first denatures and made negative by.
