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Table 1 Panel of antibodies selected in the study

From: The timing of perinatal hypoxia/ischemia events in term neonates: a retrospective autopsy study. HSPs, ORP-150 and COX2 are reliable markers to classify acute, perinatal events

Antibody against Pre--treatment Incubation time of primary antibody, temperature Concentration of primary antibody
GFAP (Dako, Copenhagen, Denmark) No pre-treatment 120 min, 20°C 1:300
TNFα (Santa Cruz, CA, USA) boiling in 0.1 M Citric Acid buffer. 120 min, 20°C 1:600
IL-1β (Santa Cruz, CA, USA) boiling in 0.25 mM EDTA buffer. 120 min, 20°C 1:4000
IL-6 (Santa Cruz, CA, USA) 5 min Proteolytic Enzyme (Dako, Copenhagen, Denmark), 20°C. 120 min, 20°C 1:2000
CD68 (Serotec, United Kingdom) 5 min Proteolytic Enzyme (Dako, Copenhagen, Denmark), 20°C. 120 min, 20°C 1:200
HSP 27 (NovaCastra, United Kingdom) boiling in 0.25 mM EDTA buffer.. 120 min, 20°C 1:20
HSP 70 (NovaCastra, United Kingdom) boiling in 0.25 mM EDTA buffer.. Overnight, 20°C 1:100
HSP 90 (NovaCastra, United Kingdom) boiling in 0.25 mM EDTA buffer.. 120 min, 20°C 1:50
COX2 (Santa Cruz, CA, USA) 5 min Proteolytic Enzyme (Dako, Copenhagen, Denmark), 20°C. 120 min, 20°C 1:100
ORP-150 (IBL, Gunma, Japan) boiling in 0.1 M Citric Acid buffer. 120 min, 20°C 1:200
β-APP (NovaCastra, United Kingdom) boiling in 0.1 M Citric Acid buffer. Overnight, 20°C 1:300
TrypH (NovaCastra, United Kingdom) boiling in 0.1 M Citric Acid buffer. Overnight, 20°C 1:50
GAP-43 (Santa Cruz, CA, USA) boiling in 0.1 M Citric Acid buffer 120 min, 20°C 1:2000
TdT enzyme (Chemicon, CA, USA) 15 min in Proteinase K (20 μg/ml), 20°C 60 min, 38°C 30 μl of TdT in 70 μl of reaction buffer (Chemicon, CA, USA)