Mouse anti ki67 polyclonal antibody

Cells were washed three times with ice-cold PBS and fixed with 4% paraformaldehyde-PBS. After 15 min of incubation with 0.1% Triton-PBS, the cells were blocked with 1% bovine serum albumin-PBS. Then, the following primary antibodies were used: (1) for hNSCs characterization, mouse anti-nestin polyclonal (1:1000, Santa Cruze, USA); (2) for hNSCs differentiation, the primary antibodies including Rabbit anti-glial fibrillary acidic protein (GFAP) (1:1000, Santa Cruze, USA), Rabbit anti-oligophrenin-4 (O4) (1:1000, Santa Cruze, USA), and mouse anti-Tuj-1(1:1000, Santa Cruze, USA) were used; (3) for hGSCs characterization, the hGSCs sphere were then incubated with mouse anti-human CD133 polyclonal antibody and mouse anti-nestin polyclonal (1:1000, Santa Cruze, USA); (4) for detection of the expressions of BMPs signaling molecules, mouse anti-human BMPR-IA, BMPR-IB and BMPR-II (Abcam, UK) were used, as well as mouse anti-Smad1 antibody, rabbit anti-phospho-Smad1 antibody (Cell signaling, USA); and (5) for examination of the proliferation and differentiation of hGSCs, mouse anti-Ki67 polyclonal antibody and mouse anti-GFAP monoclonal antibody (Abcam, UK) were used.

Mice were euthanized after 12 days post-treatment. The immunohistochemistry assay was done as described previously [20 (link)], with some modifications. Tumor tissues were stripped and fixed with 4% paraformaldehyde. Tumor tissues were embedded in paraffin and cut into sections for immunohistochemical analysis for Ki-67 (mouse anti- Ki-67 polyclonal antibody, Abcam Co., Cambridge, UK) to detect the proliferation of the tumor. In this section, the Ki-67 positive cells rate was evaluated in photomicrographs of tumor tissues (n = 3) with 200 × magnification in four fields of the tumor.