Transcription of Methods for Protein Analysis SDS-PAGE
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Methods for Protein Analysis SDS-PAGE
Methods for Protein Analysis1. Protein Separation MethodsThe following is a quick review of some common Methods used for proteinseparation: SDS-PAGE (SDS- polyacrylamide gel electrophoresis) separates proteins mainlyon the basis of molecular weight as opposed to charge (which is swamped out by the excess of Protein -bound SDS) or folding (proteins are largely denatured inSDS). In the ideal picture, the distance migrated by the Protein in a given time isinversely related to the logarithm of its molecular weight:log (mol wt.)Distancemigratedon gelHowever, factors other than molecular weight alone ( , glycosylation or astrongly hydrophobic character) can also affect the migration of a Protein on anSDS-PAGE gel, as discussed in focussing separates proteins on the basis of their balance of acidic(negatively charged) vs. basic (positively charged) amino acid residues, whichdetermines a property known as the Protein s isoelectric point [pI], the pH atwhich the Protein s net charge becomes exactly zero.
Methods for Protein Analysis 1. Protein Separation Methods The following is a quick review of some common methods used for protein separation: SDS-PAGE (SDS-polyacrylamide gel electrophoresis) separates proteins mainly on the basis of molecular weight as opposed to charge (which is ‘swamped out’
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