Monoclonal rabbit anti gfp

Adult male brains were dissected and fixed in 4% paraformaldehyde (Electron Microscopy Sciences) for 40 minutes and labeled as described previously (Certel et al., 2010 ). The following primary antibodies were used: rabbit anti-MeCP2 (1:30, Cell Signaling Technologies), mouse anti-MeCP2 (1:500, Abcam), rat anti-CD8 (1:100, Molecular Probes), monoclonal rabbit anti-GFP (1:200, Molecular Probes), mouse nc82 (1:100) and anti-MBD-R2 (1:200) (Prestel et al., 2010 (link)). Secondary antibodies include Alexa Fluor 488-conjugated donkey anti-mouse, Alexa Fluor 594-conjugated goat anti-rabbit, Alexa Fluor 647-conjugated donkey anti-mouse, Alexa Fluor 488-conjugated goat anti-rat cross-adsorbed antibodies (Jackson ImmunoResearch Laboratories, West Grove, PA). Brain samples were mounted in a drop of Vectashield^™ (Vector Laboratories Inc, Burlingame, CA) and Images were collected on an Olympus Fluoview FV1000 laser scanning confocal mounted on an inverted IX81 microscope and processed with Image-J 1.33 (NIH) and Adobe Photoshop (Adobe, CA).

The following antibodies were used in immunohistochemistry and immunocitochemistry: anti-gephyrin Mab7a (Synaptic System Cat. No 147021), anti-VGAT rabbit or guinea pig (1:1000, Synaptic System Cat. No 131004), anti-NL2 rabbit affinity purified (1:500, Synaptic system Cat, No 129203), guinea pig anti-GABA_A γ2 subunit (1:2000^{44 (link)}), biotinylated anti-guinea pig (1:200, Vector Laboratories, Cat No BA-7000). The following primary antibodies were used in immunoprecipitation and western blot analysis: mouse monoclonal anti-FLAG M2 (Sigma Cat No F1804), mouse monoclonal anti-gephyrin 3B11 (Synaptic System Cat No 147111) and rabbit polyclonal anti-NL2 (Synaptic Systems Cat. No 129202), pS/pT-P (MPM-2, Upstate Biotechnology Cat No 05-368), high affinity rat monoclonal anti-HA 3F10 (Roche), anti-GFP rabbit monoclonal (Life Technology, Cat No G10362). Validation of antibodies used in these assays can be found on the respective manufacturers’ websites.