Protein Determination by Western Blotting or Immunoblot

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Western blotting, also known as immunoblot, is a chromatography-based technique in which quantitative detection of proteins can be performed based on their molecular weight. This is often performed after the SDS-PAGE technique. It is used to identify, analyze and quantify proteins from a protein mixture. The major steps include 1. Separation of proteins (e.g., SDS-PAGE), 2. Transfer onto a solid support, 3. Utilizing primary and secondary antibodies for visualization. Western blotting can be used qualitatively and quantitatively. In this chapter, both of these techniques are explained in detail.