Free floating sections of parahippocampal gyrus were incubated with a mixture of GMF monoclonal antibody and polyclonal iNOS or polyclonal NF-κB p65 antibodies for overnight at 4°C (Thangavel et al., 2013 (link)). Then the sections were incubated for 1 h at RT with the appropriate secondary antibodies. GMF was visualized following labeling with goat anti-mouse IgG conjugated with green fluorescent dye Alexa Fluor 488, and iNOS or NF-κB were visualized post labeling with goat anti-rabbit IgG conjugated with red fluorescent dye Alexa Fluor 568. The sections were rinsed and cover-slipped with Fluorogel and the images were acquired on a Nikon (DIAPHOT) fluorescence microscope.
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