Biochemistry 2280A Chapter Notes - Chapter 5: Nuclear Magnetic Resonance, Two-Dimensional Gel Electrophoresis, Affinity Chromatography
20 Sep 2016
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In order to study proteins we need large amounts. Isolation: proteins are isolated from laboratory cells that have been genetically modified to produce more of the protein of interest. How protein analyze to determine amino acid sequence. Proteins can be purified from cells or tissues. First step in purification procedure: break open cells, release contents. Cell homogenate (extract): the contents that comes out of the cell. Initial fractionation: separates class of molecules of interest. Isolate desire protein: through series of chromatography steps. Chromatography: use different materials to separate individual components of a complex mixture, into fractions. Fractions examined to determine which contains protein of interest. Affinity chromatography: separates polypeptides on basis of ability to bind to particular molecule. Ex. large amounts of antibodies recognize protein available, they can bind to matrix of a chromatography column and used to help extract protein from mixture. Used to isolate proteins that interact physically with protein being studied.
