Chapter 2 - One-dimensional PAA-gel Electrophoretic Techniques to Separate Functional and Denatured Proteins

Abstract

This chapter describes one-dimensional polyacrylamide–gel (PAA–gel) electrophoretic techniques to separate functional and denatured proteins. The high resolution of biologically active or denatured proteins achievable with modern electrophoretic techniques is inseparably bound to the introduction of PAA as a supporting medium. PAA-gel electrophoresis first became a routine method in protein laboratories when Ornstein and Davis suggested separating human serum proteins by the use of three different buffer systems, two of which are present in two different concentration PAA gels. PAA-gel electrophoresis is usually performed with gel columns approximately the size of a pencil, but flat gels are also used, as are 5 μl glass capillaries in specialized micro-methods and 20 ml gels in sophisticated macro methods. The chapter discusses the structure and physicochemical properties of PAA-gels.