Protein Blotting by the Semidry Method

Abstract

Protein blotting, also known as Western blotting, refers to the transfer of proteins to an immobilizing membrane. The most widely used blotting method is the electrophoretic transfer of resolved proteins from a polyacrylamide gel to a nitrocellulose or polyvinylidene difluoride (PVDF) sheet and is often referred to as ``Western blotting.'' Electrophoretic transfer uses the driving force of an electric field to elute proteins from gels and to immobilize them on a matrix. This method is fast, efficient and maintains the high resolution of the protein pattern (1). There are currently two main configurations of electroblotting apparatus: (1) tanks of buffer with vertically placed wire (see Chapter 58) or plate electrodes and (2) semidry transfer with flat-plate electrodes.