LIFE 212 Lecture Notes - Electric Field, Bradford Protein Assay, Arginine
30 Mar 2014
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Document Summary
The movement of charged molecules in an electric field. Molecules separate in a porous matrix (gel) according to their charge, size and shape. Types of protein separation: native gel electrophoresis: proteins separate by size, charge, and conformation. Isoelectric focusing: proteins separate by their isoelectric point (pi) in a ph gradient pi: the ph at which protein net charge is zero: sds denatured gels: separate proteins according to size (mw) Sds: breaks noncovalent bonds, coats with ve charge. Migration of denature proteins: inversely proportional to their mw: 2d gels: 2 & 3. Separation of partially purified potato tyrosinase on sds denaturing gel electrophoresis. A standard curve is made relating the logarithm of standard protein mw size markers (y- axis) with the distance they migrate on the gel. This curve is used to determine the size (mw) of unknown proteins by measuring the distance they migrate on the gel and the line of best fit.
