CHE 350 Chapter Notes - Chapter 5: Two-Dimensional Gel Electrophoresis, Affinity Chromatography, Capillary Electrophoresis
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To p(cid:396)e(cid:448)e(cid:374)t da(cid:373)age (cid:449)he(cid:374) it"s out: keep ph stable with buffers. If you work with a ph at the protei(cid:374)"s pi, it (cid:449)o(cid:374)"t (cid:271)e (cid:448)e(cid:396)y solu(cid:271)le a(cid:374)d it (cid:449)ill be harder to work with. Salting out: salting in: when salt is added, solubility of a protein at low ion concentrations increases. Then salting out: even more salt is added, solubility of protein decreases. Ion exchange chromatography: anions bind to cationic groups on anion exchangers, and cations bind to anionic groups on cation exchangers. Proteins that bear both positive and negative charges can bind to both. The binding affinity of a particular protein depends on the presence of other ions that compete with the protein for binding to the ion exchanger and on the ph of the solution. Proteins with relatively low affinities for the ion exchanger move through the column faster than proteins that bind with higher affinities. Electrophoresis: migration of ions in an electric field.
