Pin1 antibody (MAB2294, R&D Systems) or normal mouse immunoglobulin G (IgG) (SC-2025, Santa Cruz) was immobilized on Protein G Sepharose Fast Flow Beads (P3296, Sigma-Aldrich). Beads were equilibrated in PBS for 1 hour with multiple buffer changes. Antibody was added at 4 μg per 20-μl bead slurry in PBS and allowed to bind overnight at 4°C. Beads were washed three times with fresh 0.2 M sodium borate. Freshly made dimethyl pimelimidate dihydrochloride was then used at 5.5 mg/ml in 0.2 M sodium borate for 40 min at RT to cross-link the antibody to the beads. Beads were rinsed and incubated for 2 hours at RT with 0.2 M ethanolamine. Beads were washed three times with PBS and stored up to 4 weeks.

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