In western Blotting technique the protein are transferred in nitrocellulose or nylon membrane The various steps of this techniques are described below.
(1) Proteins band separated to polyacrylamide gel electrophoresis.
(2) The protein bands are transferred onto nitrocelluolose or nylonmembrane initially this was achieved a capillary movement of buffer similar to southern blotting but now a days it is usually done by electrophoresis. Electrophoresis technique has been used in southern and nothern hybridization as well as buffer of low ionic strength and nylon membrane (since nucleic acid bind to nitrocellulose membrane only under condition of high ionic strength are used). The electrophoretic blotting both of proteins and nucleic acids is much faster and more efficient than capillary blotting.
(3) The specific protein are identified in two ways.
(1) for the specific antibodies used as a probes for detection of a specific anitgen.
(2) For the identification of glycoprotein lectin are used as a probes.