Keep MonoMac6 cells (5×10^5 cells/mL) on ice in assay buffer.
{"action": "keep", "output": "", "reagent": ["MonoMac6 cells"], "temperature": ["on ice"]}
Add assay buffer with 100 ng/mL MIF (final concentration: 50 ng/mL) Ca2+/Mg2+ (final concentration: 1 mM), incubate at 37°C for 30 min. #####%%%%% output "samples" are used in line6
{"action": "add", "output": "samples", "reagent": ["assay buffer"]}
Withdraw samples at different times, .
{"action": "withdraw", "output": "", "reagent": ["samples"], "time": ["different times"]}
add to 4% PFA (final concentration: 2%) #####%%%%% output "PBS" are used in line10
{"action": "add", "output": "PBS", "container": [""], "reagent": ["4% PFA"], "volume": [""]}
Wash cells with PBS, incubate with monoclonal antibody 327C (10 µg/mL) for 30 min. #####%%%%% output "PBS" are used in line12
{"action": "incubate", "output": "PBS", "reagent": ["PBS"], "time": ["30 min"], "temperature": [""]}
Wash cells with PBS, incubate with anti-mouse-IgG FITC conjugate for 30 min.
{"action": "incubate", "output": "", "reagent": ["PBS"], "time": ["30 min"], "temperature": [""]}
If staining is not clear, repeat the washing steps.
{"action": "repeat", "output": "", "reagent": [""]}
If the results are inconsistent, repeat the measurement steps. #####%%%%% output "the measurement steps" are not used
{"action": "repeat", "output": "the measurement steps", "reagent": [""]}