Enzyme-linked immunosorbant assays (ELISA)
Rats (N = 8 per experimental group) were injected with GTN (10 mg/kg i.p.) or vehicle and then killed with a lethal dose of anaesthetic 4 h after treatment. Their brains were immediately chopped into parts; brain areas of interest were dissected out and frozen at −80 °C until further processing.
Blood was drawn from the vena cava and centrifuged at 3000 g for 10 min at 4 °C.
ADMA levels (ng/mg proteins or nmol/ml) were quantified by ELISA kit (Antibodies Online) according to the manufacturer’s instructions.