Protein Electrophoresis Clinical Chemistry Review 89
Rate of migration Depends on size, shape, & charge of molecule
Support medium Cellulose acetate or agarose
Buffer Barbital buffer, pH 8.6
Stains Ponceau S, amido blue, bromphenol blue, Coomassie brilliant blue
Charge At pH 8.6, proteins are negatively charged & move toward anode
Order of migration (fastest to slowest) Albumin, alpha-1 globulin, alpha-2 globulin, beta globulin, gamma globulin
Largest fraction Albumin
Electroendosmosis Buffer flow toward cathode. Causes gamma region to be cathodic to point of application
Urine Must be concentrated first because of low protein concentration. Bence Jones proteins migrate
to gamma region in urine electrophoresis
CSF Must be concentrated first because of low protein concentration. CSF has a prealbumin band