Sc 21870

Immunohistochemistry was used to detect the expression of JAK2 and STAT3 proteins in brain tissue. After rat brains were removed, serial coronal sections of the entire hippocampus were cut using a sliding microtome. The sections from each brain were embedded in paraffin sections, de-waxed, hydrated and placed in an EDTA-containing antigen repair buffer (pH 9.0) to detect the immunostaining of the brain sections. Antigen retrieval was performed in a microwave oven. The slices were placed in a 3% hydrogen peroxide solution to block endogenous peroxidase and then blocked with a 5% bovine serum albumin solution at room temperature for 30 min. The sections were then incubated with goat polyclonal anti-phospho-JAK2 (Tyr 1007/1008) (SC-21870 from Santa Cruz Biotechnology, Inc.) and mouse monoclonal anti-phospho-STAT3 (B-7; Tyr 705) (SC-8059 from Santa Cruz Biotechnology, Inc.) at 4 °C overnight, followed by incubation with secondary antibodies for 50 min at room temperature. The slices were washed for 15 min and DAB color-developing solutions were used. The slides were observed under a microscope and the time was recorded. Finally, positive staining was indicated by the spots. The cells were then washed with tap water. Each slice was dehydrated and sealed after the nuclei were stained with hematoxylin.