Rabbit polyclonal anti sox2

Cells were fixed with 4% paraformaldehyde in PBS, pH 7.4 at the end of proliferation and differentiation stages. Immunocytochemistry assays were performed following standard protocols [57 (link)]. Briefly, cells were permeabilized and blocked for 1 hour with 0.3% Triton X-100 and 10% normal goat serum (NGS) in PBS. Cells were incubated overnight at 4°C with the following primary antibodies diluted in PBS containing 10% NGS: rabbit polyclonal anti-β-III Tubulin (1:2000, Covance); rabbit polyclonal anti-glial fibrillary acidic protein (GFAP; 1:2000, DAKO); mouse monoclonal antibody anti-microtubule associated protein 2 (MAP2; 1:500, Chemicon); mouse monoclonal anti-Nestin (1:100, Developmental Studies Hybridoma Bank); rabbit anti-Nestin (1:1000, Covance); rabbit polyclonal anti-TH (1:1000; Pel-freez); rabbit polyclonal anti-Sox2 (1:200; Millipore). Appropriate Alexa-Fluor secondary antibodies were used diluted in PBS (1:1000; Molecular Probes) containing 10% NGS. Nuclei were stained with Hoechst 33258 (1 ng/mL; Sigma). Immunostainings were analyzed with an epifluorescence microscope (Nikon, Eclipse TE2000-U) and photographed with a Nikon digital camera (DMX1200 F). Negative controls were performed in the absence of primary antibodies and showed no unspecific staining.