Rabbit igg bs 0295p

Immunohistochemical staining of TG and spinal trigeminal nucleus caudalis was done following our previously published work [13 (link)]. Briefly, the sections from TG were stained with primary antibody cleav Casp3 and secondary antibody (Table 3). The spinal trigeminal nucleus caudalis sections were stained with primary antibodies C-Fos and GFAP followed by secondary antibodies (Table 3). Nuclear staining was done using 4′,6-Diamidino-2-phenylindole dihydrochloride (DAPI, 1:100, Nacalai Tesque, Inc., Kyoto, Japan). Isotypes (goat IgG bs-0294P, and rabbit IgG bs-0295P, Bioss Antibodies, Boston, MA, USA) and only secondary antibodies were used as positive and negative control. Images were observed and acquired by confocal laser-scanning microscope (LSM 700, Carl Zeiss, Oberkochen, Germany).

Six to 8 microns-thick frozen sections of trigeminal ganglion were fixed in 4% paraformaldehyde, permeabilized with 0.1% Triton and subsequently blocked with Blocking One Histo (Nacalai Tesque, Kyoto, Japan). Sections were incubated with primary antibodies, rabbit anti-glutamine synthetase (1:1000; ab49873, Abcam, Cambridge, UK) and goat anti-GFAP (1:500; ab53554, Abcam), overnight at 4 °C. Sections were incubated with secondary antibodies for 2 h at room temperature using donkey anti-goat IgG Alexa Fluor 488 (1:200; ab150129, Abcam) and donkey anti-rabbit Alexa Fluor 555 (1:200; ab 150074, Abcam). For nuclear staining, 4’, 6-Diamidino-2-phenylindole dihydrochloride (DAPI, 1:100, Nacalai Tesque, Inc., Kyoto, Japan) was used for 15 min at room temperature, followed by mounting with Aqua-Poly/Mount (Polysciences, Inc., Warrington, PA, USA). Isotypes (goat IgG bs-0294P, and rabbit IgG bs-0295P, Bioss Antibodies, Boston, MA, USA) and only secondary antibodies were used as positive and negative control. Images were observed and acquired by confocal laser-scanning microscope (LSM 700, Carl Zeiss, Oberkochen, Germany).