Immunoprecipitation of mCherry-HSP90

RFP Selector (NanoTag #N0410) resins were equilibrated with lysis buffer to prepare the resin. Cell lysates were then added and incubated with the resins at 4°C with head over tail rotation for 90 min. Following incubation, resins were washed twice with lysis buffer and once with PBS before elution with 2 × sample buffer and incubation at 95°C for 5 min. Eluents were then run on a 12.5% SDS-PAGE. For SILAC samples, heavy and light replicates were immunoprecipitated separately, before combined and separated by SDS gel electrophoresis.