A Dot Blot is a simple and quick assay that may be employed to determine if your antibodies and detection system are effective. Dot Blot may also be used to determine appropriate starting concentration of primary antibody for Western blot.
This protocol is intended to provide a set of initial conditions for analysis of cell lysates samples by Western blot. Further optimization may be required for individual samples or analytes. Follow manufacturer's protocols for specific reagents when applicable.
R&D Systems Quality Control laboratories use these Western blotting and immunostaining protocols to show that our polyclonal and monoclonal antibodies are specific for the proteins they were raised against and to determine the sensitivity of the antibody for its antigen.