Co-immunoprecipitation was performed as previously described (Qin et al., 2014 (link)). Protein A/G plus agarose beads (Santa Cruz) first pre-cleared with whole cell lysates, which were then incubated with anti-β-arrestin1 (no: 32097, abcam) under gender agitation overnight at 4°C. Next day the Protein A/G plus was mixed with the lysates and the mixture was incubated for 3 h at 4°C rotating. Following special centrifugation, the precipitation was washed three times with whole cell lysates and boiled in 1x SDS-loading buffer. Finally, the samples were subjected to normal Western Blot procedures.
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