Enzyme Kinetics IV - Experiment 15: Isolation,...

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: , , Experiment 15 Isolation Purification and Kinetics of . E coli Alkaline Phosphatase Part 4 Biochemistry 332L - - 2 26 09 This week you will be learning to pour polyacrylamide gels for ( ). , Polyacrylamide Gel Electrophoresis PAGE As we learned last semester these gels are formed by creating multiple bonds between molecules of (“ - ”) ”. acrylamide cross linking to form a matrix The matrix is the web of - , polymer strands that results from the cross linking process through which . protein or DNA molecules will migrate when pulled by an electric field All , else being equal smaller molecules will migrate through the matrix faster . than larger ones Few undergraduate labs teach students how to pour polyacrylamide , gels because prior to polymerization acrylamide is a potent and deadly . , , . neurotoxin For this week s lab then safety is our number one concern . Goggles and Gloves are mandatory I will do most of the handling of the . acrylamide . There are two types of polyacrylamide typically used with proteins The - , first is the SDS PAGE gel which contains the detergent sodium dodecyl ( ). sulfate SDS This type of gel is used to run proteins in their denatured ; state the open polypeptide chain becomes associated with the SDS , detergent and acts as though it were simply a long chain molecule . exhibiting no secondary or tertiary structure This is desirable when the . , molecular weight of the protein is to be determined In general natural ( proteins have molecular weights proportional to their chain lengths although , different amino acid residues have different weights the composition of , proteins does not vary widely that is the average proportion of the
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Enzyme Kinetics IV - Experiment 15: Isolation,...

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