SDS PAGE Electrophoresis

SDS PAGE Electrophoresis - Protein Marker Molecular Weight...

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Protein Marker Molecular Weight (MW) Log MW Protein Migration (cm) Buffer front (cm) R f Myosine, Porcine 200,000 5.301 0.9 6.0 .15 β Galactosidase, E. Coli 116,000 5.064 1.8 6.0 .30 Phosphorylase B, Rabbit Muscle 97,000 4.987 2.0 6.0 .33 Albumin, Bovine 66,000 4.820 2.9 6.0 .48 Ovalbumin, Chicken 45,000 4.653 4.3 6.0 .72 Carbonic Anhydrase, Bovine 29,000 4.462 5.3 6.0 .88 Migration (cm) Buffer front (cm) R f Log MW MW Milk Protein 1 4.85 6.0 .81 Milk Protein 2 5.30 6.0 .88 Tables and Graphs Determining the Unknown
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Questions 1. How is it possible to use electrophoresis, a separation technique depending on molecular charge, to separate proteins based on molecular weight? A porous gel allows small molecules to move freely while retarding larger molecules. Smaller molecules, then, move further down the gel than do the larger molecules, making a clear distinction based on molecular weight. However, this works only if all the molecules have the same charge so that they move in the same direction. 2. Why do we generally use polyacrylamide gel instead of agarose gel for protein separations? Agarose gel is used when attempting to separate larger molecules like nucleic acids and large proteins and protein complexes because its pores are larger. The pores of
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SDS PAGE Electrophoresis - Protein Marker Molecular Weight...

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