BIO SCI 98 Lecture Notes - Lecture 4: Affinity Chromatography, Column Chromatography, Centrifugation
3 Aug 2018
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Document Summary
Active site of an enzyme: a small cleft in the enzyme that has a defined 3d structure. The standard conditions of an enzyme catalyzed reaction. Homogenization: prepare cell by cracking it open and gathering the free extract. Centrifugation: putting it in a machine that quickly spins the mixture so that the heavy structures move to the bottom; this will remove membranes, nuclei, large organelles aka mitochondria. Salting-in and salting-out: based on protein solubility. Salting in: at low concentrations of salt, the protein"s solubility increases. Salting out: at high concentrations of salt, the protein"s solubility decreases. Salting out separates desired proteins by adding salt to a solution that causes the water molecules to be gradually exhausted from the surrounding of the desired molecules that are polar in water. This causes the desired proteins to come together and settle down. Column chromatography: an impure sample is loaded onto a column filled with silica.
