Usage of a semi-dry electrophoretic apparatus is a fast and cost effective way to transfer proteins from polyacrylamide gels to membranes for immunoblotting. However, its efficiency is usually limited to small and mid-range molecular weight proteins, and the time-consuming wet transfer procedure is usually the preferred method for high molecular weight proteins. Here we describe a protocol for the efficient semi-dry transfer of proteins of diverse sizes (ranging from 15 to 296 kDa) resolved in a gradient SDS-PolyAcrylamide Gel Electrophoresis (PAGE) gel. This protocol was successfully employed to transfer and detect the small protein Rps22 (15 KDa), the high molecular weight protein Gcn1 (296 kDa), among others. This protocol is also suitable for the quantitative detection of phosphorylated proteins such as phosphorylated eIF2 alpha.