Rabbit anti gfap z0334

Poly‐d‐Lysine (PDL) (P0899), DNase 1 type IV, Triton X‐100, FITC‐NHS, Rhodamine 6G (Rh6G), fluorescein sodium salt (FITC), and 1,1′‐Dioctadecyl‐3,3,3′,3′‐tetramethylindocarbocyanine perchlorate were purchased from Sigma‐Aldrich. Alexa‐Fluor 488 goat anti‐mouse, 4′,6‐diamidino‐2‐phenylindole, paraformaldehyde (PFA, 7 230 681), goat serum, Dulbecco's modified eagle medium (DMEM), penicillin/streptomycin (Pen/Strep), B27 supplement (17 504 001), GlutaMAX, neurobasal medium, Fluoromount‐GTM mounting medium (00‐4958‐02), fetal bovine serum (FBS), CMFDA, and YO‐PRO‐1 iodide were purchased from Life Technologies. Mouse anti‐Tuj1 antibody (801 202) was purchased from Biolegend. Rabbit anti‐GFAP (Z0334) was obtained from DAKO. Papain suspension was purchased from Worthington. Cell strainer (70 µm) was obtained from BD, Biosciences, USA.

TNFα, LTα₁β₂, IL-1β, B-cell activating factor (BAFF), CD40L, and LIGHT were obtained from PeproTech. APQ [NF-κB activation inhibitor, 6-amino-4-(4-phenoxyphenylethylamino)quinazoline] was from EMD Chemicals and dissolved in DMSO. Antibodies against LTβR(M-110), Calretinin(N-18), Doublecortin (DCX, C-18), Myelin basic protein (MBP, D-18), Sox2(Y-17), p65(C-20), GAPDH and Lamin A/C(636) were obtained from Santa Cruz Biotechnology. RelB and p100/p52 were from Cell Signal Technology. Mouse anti-NeuN antibody was from Millipore. Chicken anti-glial fibrillary acidic protein (GFAP), anti-Neuron-specific class III β-tubulin (Tuj1), and anti-Nestin antibodies were from Aves. Rabbit anti-GFAP (Z0334) was from Dako. All other reagents were from Sigma.

For whole mount and cryosection immunostaining, embryos were fixed in 4% paraformaldehyde (PFA) at 4°C overnight at different stages (between 11 to 24 hpf). Embryos were blocked in 10% normal goat serum (Sigma-Aldrich, St Louis, MO, USA) for 2 hours at room temperature. The following primary antibodies were used in this study: mouse-anti-ZO-1 (339111; Zymed Laboratories, South San Francisco, CA, USA) at 1:300; rabbit-anti-aPKC (C-20; Santa Cruz Biotechnology, Dallas, TX, USA) at 1:500; rabbit-anti-GFAP (Z0334; DakoCytomation, Glostrup, Denmark) at 1:500; mouse-anti-MF-20 (Developmental Studies Hybridoma Bank, Iowa City, IA, USA) at 1:50; mouse-anti-acetylated-tubulin (T6793; Sigma-Aldrich); and rabbit-anti-phospho-histone H3 (Upstate Biotechnology, Lake Placid, NY, USA) at 1:200, diluted in 2.5% normal goat serum (Sigma-Aldrich). For secondary antibodies, anti-rabbit and anti-mouse Alexa 488, Alexa 568 and Alexa 633 (Molecular Probes, Eugene, OR, USA) were used at 1:800 in 2.5% normal goat serum. Sections were cut every 14 to 16 μm on a Leica 5100 or Cryo-Star HM 560 MV Micron microtomes.