The incubation step in western blotting
Western blotting is one of the most reliable procedures for identifying a protein(s) in a mixture of other proteins. It involves eight steps to separate and visualize the protein of interest, they include; Sample preparation Gel electrophoresis. Protein/membrane transfer. Blocking Incubation with primary antibody. Secondary incubation Detection. Analysis. The incubation step is one of the most crucial procedures in western blotting. It occurs after the blocking stage and before final detection and visualization. Ensuring thoroughness during this step is vital to the success of the procedure. In this blog, you will learn about the incubation phase and the required criteria for successful immunoblotting. Incubation is a two-part process that involves the use of primary and secondary antibodies. In simple terms, the primary antibodies bind to the protein of interest. To detect the proteins, the secondary antibodies need to bind to the primary ones. Some factors that affect the quali...