Collect, at ~1-2 x 10^6 cells/ml in RPMI 1640 media (no FBS).
{"action": "collect", "output": "", "container": ["RPMI 1640 media (no FBS)"]}
Split samples, treat one with 100 ng/ml PTX, .
{"action": "treat", "output": "", "concentration": ["100 ng/ml PTX"]}
rotate at 37°C for 3-4 h ()
{"action": "rotate", "output": "", "temperature": ["37\u00b0C"], "time": ["3-4 h"]}
Protect tubes from light, .
 {"action": "protect", "output": "", "container": ["tubes"]}
rotate for 30 min at 37°C
{"action": "rotate", "output": "", "time": ["30 min"], "temperature": ["37\u00b0C"]}
If staining is incomplete, repeat the staining steps. #####%%%%% output "the staining steps" are used in line16
{"action": "repeat", "output": "the staining steps", "reagent": [""]}
Stimulate cells with 50 µl 10 µg/ml SDF-1, record responses for 3 min, then stimulate with 50 µl 20 µM ionomycin.
{"action": "induce", "output": "", "concentration": ["50 \u00b5l 10"], "reagent": ["ionomycin"], "time": ["3 min"]}
If responses are not clear, repeat the stimulation . #####%%%%% output "the stimulation" are not used
{"action": "repeat", "output": "the stimulation", "reagent": ["the staining steps"]}
Note: Replace SDF-1 with other chemokines as desired, determine optimal dose.
{"action": "note", "output": "", "reagent": ["SDF-1", "other chemokines"], "bool": [""]}