《Tricine-Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis》.pdf

ANALYTICAL BIOCHEMISTRY 166. 368-379 (I 987) Tricine-Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis for the Separation of Proteins in the Range from 1 to 100 kDa HERMANN SCH.GGER AND GEBHARD VON JAGOW Received February 25, 1987 A discontinuous sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS) system for the separation of proteins in the range from I to 100 kDa is described. Tricine, used asthe trailing ion, allows a resolution of small proteins at lower acrylamide concentrations than in glycine-SDSsystems. A superior resolution of proteins, especially in the range between 5 and 20 kDa. is achieved without the necessityto use urea. Proteins above 30 kDa are already destacked within the sample gel. Thus a smooth passageof these proteins from sample to separating gel is warranted and overloading effectsare reduced. This is of special importance when large amounts of protein are to be loaded onto preparative gels. The omission of glycine and urea prevents disturbances which might occur in the course of subsequent amino acid sequencing. ic 19X7 Academic Press. Inc KEY WORDS: SDS: proteins; peptides. The SDS’ system, in part already of Davis (7) and Ornstein (8) was chosen as published ( 1), was developed becausea sepa- the basisfor the method described. ration of the I 1 protein subunits of the bcr The stacking limit for large proteins in the complex from beef heart was not