{"action": "harvest", "device": ["centrifugation"], "force": ["2000 RPM"], "output": "", "time": ["5 min"]}
{"action": "homogenize", "output": "", "reagent": ["cell lysates"]}
{"action": "centrifuge", "force": ["2500 RPM"], "output": "", "time": ["5 min"]}
{"action": "transfer", "container": ["fresh centrifuge tubes"], "output": ""}
{"action": "add", "container": ["each sample"], "output": "residual insoluble proteins", "reagent": [""]}
{"action": "spin", "device": ["ultracentrifugation"], "force": ["55000 RPM"], "output": "Hypotonic Buffer", "reagent": ["residual insoluble proteins"], "time": [""]}
{"action": "transfer", "container": ["fresh centrifuge tubes"], "output": "", "reagent": ["supernatants"]}
{"action": "rinse", "output": "use", "reagent": ["Hypotonic Buffer"]}
{"action": "take", "output": "Hypotonic Buffer", "volume": ["cell lysates"]}
{"action": "dilute", "output": "antibody", "reagent": ["Hypotonic Buffer"]}
{"action": "add", "output": "polyclonal anti-MEKK1", "reagent": ["Protein A beads", "antibody"]}
{"action": "add", "container": ["the lysates"], "output": "", "reagent": ["polyclonal anti-MEKK1"]}
{"action": "rinse", "output": "", "volume": [""]}
{"action": "perform", "device": [""], "output": "Hypotonic Buffer"}
{"action": "rinse", "output": "", "reagent": ["Hypotonic Buffer"]}
{"action": "use", "output": "M", "reagent": ["", ""]}
{"action": "incubate", "concentration": [""], "container": ["Kinase Assay Buffer"], "output": "", "reagent": ["M"], "temperature": ["30 \u00b0C"]}