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Cells were lysed with MCLB buffer without phosphatase inhibitors followed by debris removal via centrifugation and immunoprecipitation with pre-equilibrated anti-HA-beads overnight. Then, beads were washed with MCLB buffer for three times and incubated with Lambda Protein Phosphatase (PPase, New England Biolabs, Ipswich, MA) for 1 hr according to manufacturer’s instructions, prior to elution with 4x laemmli buffer and boiling at 95°C for 5 min. Samples were then analyzed by SDS-PAGE and immunoblotting.
Copyright and License information: ©2017, Jung et al
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