N terminal

Ten micron (μm) thick serial brain sections were prepared starting from the needle entry site to 1.0 mm anterior and posterior to injection site. Tracing of GFP-labelled human OBNSCs following engraftment was conducted using Anti-GFP (1:2000, rabbit, N-Terminal, Sigma-Aldrich).
Visualization of the primary antibodies was done using Biotin-conjugated goat anti rabbit secondary antibody labeled with poly-horseradish peroxidase (HRP) conjugate and metal enhanced DAB to develop the signal (brown color).

At the end of behavioral testing, the animals were anesthetized, and the SC tissues cryoprotected in 30% sucrose were cronally sectioned, and stained with Anti-GFP (1:2000, rabbit, N-Terminal, Sigma-Aldrich) and cell type-specific markers. The following primary antibodies were used: anti-Nestin (1:200, rabbit, Sigma-Aldrich) for detection of undifferentiated NSCs; anti-MAP2 (1:200, rabbit, Sigma-Aldrich) for detection of mature neurons; anti-beta-tubulin ІІІ (1:100, rabbit, Sigma-Aldrich) for detection of immature neurons; anti-O4 (1:100, mouse, monoclonal, Sigma-Aldrich) for detection of immature oligodendrocytea; and anti-GFAP (1:400, mouse, clone G-A-5, Sigma-Aldrich) for detection of astrocytes. Fluorescine iso-thiocyanate, (anti-rabbit, anti-mouse, anti-chicken, Sigma-Aldrich), tetra methyl rhodamyne iso-thiocyanate (anti-rabbit, Sigma-Aldrich) and Phycoerythrin (anti-mouse, anti-goat, Sigma-Aldrich) were applied, and then the SC tissues were examined under a fluorescent microscope.