Fitc conjugated rabbit anti rat igg

For double-immunofluorescence staining, tissue sections were processed as described elsewhere (Park et al., 2007) . Briefly, sections were collected in PBS and sections were incubated in 0.2% Triton X-100 for 30 min and rinsed three times with 0.5% BSA. The sections were then incubated with the indicated primary antibodies (rabbit anti-TonEBP (Miyakawa et al., 1999) , 1:1000; mouse anti-OX-42, 1:400 (Serotec, UK); mouse anti-NeuN, 1:1000 (Millipore, US); mouse anti-GFAP 1:1500 (Abcam, US)) overnight at 4 °C. After washing in PBS, the sections were incubated with secondary antibodies (FITC-conjugated rabbit anti-rat IgG, 1:1000; Texas Red-conjugated donkey anti-goat IgG, 1:1000 (Abcam, USA)) for 1 h at room temperature. Floating sections were mounted on gelatin-coated slides and dried for 1 h at room temperature. Slides were mounted by adding Vectashield medium (Vector Laboratories). Slides were imaged by using an LSM 700 confocal laser scanning microscope (Carl Zeiss, Germany).