for 5 min to pellet insoluble material and then incubated with Protein-A Sepharose for 30 min as a preclearing step. Following a second spin at 10,000 g for 5 min, the lysate was then treated with antibodies against the target proteins for 90 min after which Protein-A Sepharose was added to capture the immune complexes. The Protein-A Sepharose was subjected to multiple washes before desiccation in a speed vac and then analysis by western blotting or by mass spectrometry.
