Western Blotting

Western blotting is used to transfer proteins from polyacrylamide gels (PAGs) to the membranes, making its identification and determination more convenient. For this, PAG and membrane are sandwiched together between two parallel electrodes. On passage of current, proteins transfer from the gels to the membranes. This chapter outlines various components, merits and demerits of western blotting technique. Most commonly used blotting membranes, namely, nitrocellulose membranes, polyvinylidene difluoride (PVDF), and nylon, are also compared for their properties, merits and demerits in this section. Further, visualization and detection of protein markers and targeted proteins over the membrane can be achieved using various protocols. Ponceau S or Congo red dyes are frequently employed for the detection of protein markers, whereas for the detection of analyte (specific protein) antigen antibody interaction, enzyme-based interaction of carbohydrate moiety reaction can be utilized. Lastly, its applications in dairy science are also outlined.