western blotting protocol Archives - Gen9 Genetics

Western blot is one of the most widely used techniques in molecular biology and proteomics. Since western blot is a multi-step protocol, variations and errors can occur at any step, reducing the reliability and reproducibility of this technique. Recent reports suggest that some key steps, such as the sample preparation method, the amount and source of the primary antibody used, as well as the normalization method used, are critical to obtaining reproducible Western blot results. Areas covered: In this review, improvements in different areas of Western blotting are summarized, including protein blotting and antibody validation. The review looks at the most advanced Western Blot techniques available and highlights the relationship between next-generation Western Blot techniques and their clinical relevance. Expert Comment: Significant improvements have been made over the past decade in creating more sensitive, automated, and advanced techniques by optimizing various aspects of the Western blot protocol. New methods such as Single Cell Resolution Western Blot, Capillary Electrophoresis, DigiWest, Automated Microfluidic Western Blot, and Microchip Electrophoresis have been developed to reduce the potential problems associated with the Western Blot technique. Innovative developments in instrumentation and increased sensitivity for Western blots offer novel possibilities to increase the clinical implications of Western blots.