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) |