Lecture9 Notes

Lecture9 Notes - Lecture 9 Solution Characterization of...

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Lecture 9 Solution Characterization of Polymers III ANNOUNCEMENT: Review Session will be Tuesday during normal lecture time. Equation sheet and practice exam are posted Exam will cover lectures 1-9 1 Wednesday, October 20, 2010
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2 Technique Information Comments SEC HPLC Osmotic Pressure Viscosity MALDI, Mass Spec Fractionation Ultracentrifugation End Group Analysis Cryoscopy, Ebulliometry VPO SLS DLS Wednesday, October 20, 2010
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Dilute Solutions of Biomacromolecules 3 Why are biomacromolecules difficult to work with? Wednesday, October 20, 2010
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Gel Electrophoresis Electromotive force is used to push or pull the charged molecules through the matrix Molecules with a negative charge move towards the anode, and a positive charge towards the cathode At the end of the run, the smallest molecules reach the anode/cathode, the gel is stained to make the macromolecules visible Typical stains include coomassie blue, ethidium bromide, silver, or the analytes may fluoresce The distance traveled is proportional to the molecular weight, and molecular weight markers are used to “calibrate” the gel •Gel electrophoresis is used to separate macromolecules by charge •The gel is crosslinked polymer - poly(acrylamide) for proteins and small nucleic acids, and agarose for larger nucleic acids (>100 bp) 4 Wednesday, October 20, 2010
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SDS-PAGE SDS-PAGE: Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis Used to separate proteins according to electrophoretic mobility, which is a function of length, molecular weight, higher order protein folding, post- translational modifications etc
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