BIOL 200 Lecture Notes - Lecture 37: Chromogenic, Sodium Chloride, Trypsin
22 Jun 2016
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Document Summary
Liquid chromatography liquid chromatography: useful for isolation of large amounts of proteins. Principle: columns filled with polymer beads slow down all proteins down their path; size of meshwork within the beads is calculated in a way that proteins have the probability to go into the beads or bypass the beads. Based on size/mass (gel filtration), charge (ion exchange), and binding affinity. Any protein bigger than the pore size of polymer breads will not be separated; separation comes with smaller proteins. Anion exchange: beads bind to negatively charged proteins; also cation exchange layer sample on column, positively charged gel bead collects negatively charged protein; elute negatively charged protein with salt solution (nacl) Elution with nacl: low salt = all negatively charged particles bind; medium salt = weakly charged proteins are eluted; high salt = highly charged proteins are eluted. Charge of protein depends on ph, so binding of protein can be influenced by change in buffer ph.
