The Design of a Quantitative Western Blot Experiment

Figure 1

Reliable western blot data can only be generated when the proper sample amount of protein is used. Loading too much protein leads to signal saturation in western blots, yet too little produces weak signals. Once the experimental setup and conditions are established for the assay, do not change the sample load, transfer method, transfer time, antibody dilution, antibody incubation time, or temperature in subsequent experiments as these factors may significantly change the detection signals.