07) 08 Sept 2010

07) 08 Sept 2010 - Gels are prepared using agarose a...

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Gels are prepared using agarose, a polymer extract from seaweed, or acrylamide, which polymerizes through a free radical reaction Soft gels which compress under pressure – use electric field instead Separation is then based on size and charge
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The detergent SDS is used to denature proteins and make them travel more uniformly through the gel The sulfate group dominates the charge therefore all proteins travel in the same direction SDS-PAGE = SDS polyacrylamide gel electrophoresis Separate primarily by size since charge is equalized by SDS
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Standard proteins of know MW are used for comparison Gradient gels help develop a more linear separation Interpolation is used to estimate MW of protein of interest
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Polyionic polymers, ampholines, are added to a gel In an electric field, ampholines produce a pH gradient A protein in the ampholine gradient will migrate to a pH = pI Isoelectric focusing (IEF)
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Two-dimensional gel electrophoresis combines IEF and SDS-PAGE First separate by pI Then separate by size
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Crude extracts can be separated on 2D gels Compare different conditions and find out what changes in an organism = proteomics
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Primary structure = amino acid sequence Secondary structure = local folding domains Tertiary structure = fold of an entire polypeptide chain Quaternary structure = interaction of multiple polypeptides
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This note was uploaded on 09/18/2011 for the course CHEM 3510 taught by Professor Mueser during the Spring '10 term at Toledo.

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07) 08 Sept 2010 - Gels are prepared using agarose a...

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