Rain tissue and serum enzyme-linked immunosorbent.Assay (ELISA).,,, At 0.5 1 3 and 7 days, after MCAO rats (n = 5 - 6 per group.Per time point) were deeply anesthetized with 10%, chloral hydrateAnd blood samples were collected from the abdominal.Aorta. The brains were removed and immediately dissected on.Ice. The cerebral hemisphere ipsilateral to the MCAO was dissected.Out and stored at − 80 ° C. Subsequently brain samples,,Were homogenized in PBS buffer the homogenate, was centrifuged.For 45 minutes, at 9 727 × g at 4 ° C and the, supernatant.Was collected. Concentrations of brain lysate and, serum NT3As well as brain levels of growth-associated protein 43 (GAP -.43), were quantified using a commercially available NT3 and.GAP-43 ELISA Kit (Ever Systems Biology Laboratory Sacramento,,CA, the USA). Protein concentrations were determined.Using the bicinchoninic acid method.
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