Rapid Screening of the Epidermal Growth Factor Receptor Phosphosignaling Pathway via Microplate-Based Dot Blot Assays
Figure 5
Native protein detection—primary antibody optimization. Serum-starved A431cells were stimulated with EGF for 20 min and harvested and lysates prepared as described previously. Dot blot assays were performed to optimize detection by (a) GAPDH and (b) phospho-ERK1/2 antibodies. A series of seven lysate concentrations (100–10000 ng/well) was used to assess six different dilutions of each primary antibody. All points were run in triplicate.
