{"action": "wash", "force": ["300 x g"], "output": "", "reagent": ["PBS"], "temperature": ["ice cold"], "time": ["3 min"]}
{"action": "transfer", "container": ["new microcentrifuge tube"], "output": "the clear supernatant"}
{"action": "incubate", "output": "", "temperature": ["4 \u00b0C"]}
{"action": "add", "output": "", "reagent": ["protein G sepharose beads (50% [v/] in PBS)"], "volume": ["30 \u00b5l"]}
{"action": "incubate", "output": "the immunoprecipitates", "temperature": ["4 \u00b0C"], "time": ["2-4 h"]}
{"action": "wash", "output": "the supernatant", "reagent": ["the immunoprecipitates"]}
{"action": "wash", "output": "the supernatant", "reagent": ["the immunoprecipitates"]}
{"action": "stop", "output": "the reaction", "reagent": ["SDS-PAGE loading buffer"]}
{"action": "cover", "output": "radioactive contamination", "temperature": ["95 \u00b0C"], "time": ["10 min"]}
{"action": "open", "concentration": [""], "device": ["exposure cassette"], "output": ""}
{"action": "place", "container": ["the cassette", "the wrapped gel"], "output": ""}
{"action": "place", "device": ["the phosphor-storage screen"], "output": ""}
{"action": "place", "output": "", "temperature": ["room temperature"], "time": ["30 min"]}
{"action": "take", "device": ["the phosphor-storage screen"], "output": "direct light", "volume": [""]}
{"action": "detect", "device": ["the Typhoon Trio+ system"], "output": "the screen"}
{"action": "advise", "device": ["", ""], "output": "recombinant PKC-\u03b8"}
{"action": "activate", "output": "", "reagent": ["T538", "direct light", "recombinant PKC-\u03b8"]}
{"action": "conduct", "device": [""], "output": "the kinase assay"}
{"action": "add", "output": "the kinase assay", "reagent": ["ATP-competitive PKC-\u03b8 inhibitor"]}