feb 17 - Today: Finish protein separation Begin cell...

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Today: Finish protein separation Begin cell culture techniques Protein separation techniques Isoelectric focusing o Seldom used by itself, combined with 2d electrophoresis o Separates proteins based on isoelectric point. There is a power supply. There is a Ph gradient as well. Point where they migrate then not migrate anymore. o Very high resolution system. Serum and get 40+ different bands. They can be as close as .01 PH unit apart. 2D electrophoresis o Isoelectric focusing separates them, followed by SDS gel. o Use isoelectric focusing to separate protein in one direction. o Process Run SDS is other direction. If one band consists of 3 different proteins, then 3 dots may be seen. 2D gel. We can scan the gel in an imaging system; digitizes data. o Difficult to do; needs to be done on regular basis to do well. Western Blot o Combination of Gel electrophoresis and ‘blotting’ o Try to identify a protein using an antibody probe. o Process Run gel, flip over. Transfer onto transfer paper, which is like a sponge, using a power supply. Power supply drives proteins out of gel onto paper. Superficially looks like gel. Use indirect technique to add antibody with a tag. Quantitative; you can see an actual amount.
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This note was uploaded on 04/13/2009 for the course BIOL 311 taught by Professor Vanbuskirk during the Spring '08 term at Binghamton University.

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feb 17 - Today: Finish protein separation Begin cell...

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