gel filtration column_pic - e.g. if upper limit is 80kDa,...

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Gel filtration/Size Exclusion Chromatography Resin (beads) has tiny holes that act like a molecular sieve Proteins that fit through those holes have to go on a ‘detour’ on the way down the column the smaller the protein, the more often a protein enters a hole (more likely) and the later it comes out the column (at the bottom) larger proteins will still enter holes, but not as often, therefore come out earlier upper limit : o only proteins that actually fit into the holes will be separated, all the proteins that are to large will go straight through and come out together
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Unformatted text preview: e.g. if upper limit is 80kDa, all proteins with MW > 80kDa will come out together (at the beginning) lower limit : o the lower limit is determined by probability: the smaller the protein the more likely it will enter a hole (smaller > more holes > longer way). But, at a certain size the likelihood of entering a hole diverges and small Proteins cant be separated anymore e.g. if lower limit is 20kDa, all proteins with MW < 20kDa will come out together (last), because they all have to travel the longest way...
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This note was uploaded on 02/02/2010 for the course BIOCHEM Biochem440 taught by Professor Dr.klevit during the Winter '09 term at University of Washington.

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