Table 1

Panel of antibodies selected in the study

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)

Riezzo et al. Diagnostic Pathology 2010 5:49   doi:10.1186/1746-1596-5-49