Western Blotting: A Powerful Technique for Protein Analysis

Western blotting, also known as protein immunoblotting, is a method used to detect specific proteins in a sample of tissue homogenate or extract. It uses gel electrophoresis to separate native or denatured proteins by the length of the polypeptide chains. The proteins are then transferred to a membrane, where they are stained with antibodies specific to the target protein. The antibodies are typically conjugated to an enzyme or radioactive molecule, allowing visualization of the protein bands and quantitation of protein levels in different samples.

Sample Preparation

The first step in Western Blotting is preparing the tissue or cell extract containing the proteins of interest. Common techniques include homogenizing or lysing the sample to release intracellular proteins. Protease and phosphatase inhibitors are often added to the extraction buffer to prevent degradation of proteins during preparation. The extract is then centrifuged to remove cell debris and organelle