Mouse anti ctbp1

Briefly, brain slices and N27 cells were incubated in a blocking solution (2% FBS and 0.3% Triton X-100 in PBS) for 2 h at room temperature. Brain slices were incubated with the primary antibodies in a blocking solution for 2 overnights (mouse anti-CtBP1,1:1000, BD Biosciences, cat no. 612042; mouse anti-CtBP2, 1:1000, BD Biosciences, cat no. 612044; rat anti-CD11b, 1:1000, Serotec; rabbit anti-GFAP, 1:200, DAKO; rabbit anti-NeuN, 1:500, Cell Signaling; rabbit anti-TH, 1:1000, Santa Cruz Biotechnology), while N27 cells were incubated for 1 overnight (mouse anti-CtBP1 and mouse anti-CtBP2, 1:200) at 4 °C. Afterward, tissue and cells were incubated for 2 h at room temperature with the following appropriated secondary antibodies: Alexa Fluor 594 donkey anti-mouse (Abcam), Alexa Fluor 488 donkey anti-rat (Life Technologies) and Alexa Fluor 647 donkey anti-rabbit (Life Technologies) (1:1000 for tissues and 1:200 for cells). Lastly, sections were rinsed with PBS and mounted in Fluoroshield Mounting Medium (Abcam). Images were acquired under the magnification of 40 × using a Zeiss inverted confocal microscopy (Axiobserver Z1, Zeiss).