Incubate slides in EDTA at 95°C for 50 min for antigen unmasking.
{"action": "incubate", "output": "", "reagent": ["EDTA", "antigen"], "temperature": ["95°C"], "time": ["50 min"]}
Cool slides for 20 min at room temperature, #####%%%%% output "H2O" are used in line6
{"action": "rotate", "output": "H2O", "temperature": ["room temperature"], "time": ["20 min"]}
wash in H2O.
{"action": "wash", "output": "", "reagent": ["H2O"]}
Incubate slides in 3% H2O2 for 5 min at room temperature. #####%%%%% output "TBS" are used in line10
{"action": "rotate", "output": "TBS", "reagent": ["3% H2O2"], "temperature": ["room temperature"], "time": ["5 min"]}
Wash twice in TBS.
{"action": "wash", "output": "", "reagent": ["TBS"]}
Preincubate slides with Ab mix for 20 min at room temperature. #####%%%%% output "Ab mix" are used in line14
{"action": "rotate", "output": "Ab mix", "temperature": ["room temperature"], "time": ["20 min"]}
Incubate slides overnight at 4°C with anti-γH2AX in Ab mix, . #####%%%%% output "TBS" are used in line16
{"action": "incubate", "output": "TBS", "reagent": ["anti-γH2AX", "Ab mix"], "temperature": ["4°C"]}
wash twice in TBS #####%%%%% output "secondary antibody" are used in line18
{"action": "wash", "output": "secondary antibody", "reagent": ["TBS"]}
Incubate with secondary antibody for 30 min at room temperature, . #####%%%%% output "TBS" are used in line20
{"action": "incubate", "output": "TBS", "reagent": ["secondary antibody"], "temperature": ["room temperature"], "time": ["30 min"]}
wash twice in TBS #####%%%%% output "peroxidase substrate solution" are used in line24
{"action": "wash", "output": "peroxidase substrate solution", "reagent": ["TBS"]}
If washing is incomplete, repeat the washing steps.

Incubate in peroxidase substrate solution for 5 min at room temperature. #####%%%%% output "positive (irradiated skin)" are used in line28
{"action": "incubate", "output": "positive (irradiated skin)", "reagent": ["peroxidase substrate solution"], "temperature": ["room temperature"], "time": ["5 min"]}
Dehydrate sections through graded alcohol series, mount with Eukitt.

Include positive (irradiated skin) .
{"action": "include", "output": "", "reagent": ["positive (irradiated skin)"]}
If control results are inconsistent, repeat the staining steps. #####%%%%% output "the staining steps" are not used
{"action": "repeat", "output": "the staining steps", "reagent": [""]}