Rapid Screening of the Epidermal Growth Factor Receptor Phosphosignaling Pathway via Microplate-Based Dot Blot Assays

Figure 2

The dot blot protocol. Depicted in the diagram is the assay for one representative well from a 96-well microplate. After protein binding, a blocking step is performed to reduce nonspecific Ab binding to any unoccupied region of membrane. The remaining portion of the assay is similar to ELISA except that all wash steps are performed via vacuum filtration.