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Immunoprecipitation protocols generally comprise lysing a population of cells in a lysis buffer such as RIPA buffer (1% NP-40 or Triton X-100, 1% sodium deoxycholate, 0.1% SDS, 0.15 M NaCl, 0.01 M sodium phosphate at pH 7.2, 1% Trasylol) supplemented with protein phosphatase and/or protease inhibitors (e.g., EDTA, PMSF, aprotinin, sodium vanadate), adding the antibody of interest to the cell lysate, incubating for a period of time (e.g., 1-4 hours) at 4 C., adding protein A and/or protein G sepharose beads to the cell lysate, incubating for about an hour or more at 4 C., washing the beads in lysis buffer and resuspending the beads in SDS/sample buffer

doi: 10.1186/s12934-017-0646-4

EMBO Rep 24 , EMBR202255556 (2023)