Ab2033

For Immunostaining, brain sections were permeabilized and blocked with PBS containing 10% normal goat serum, 10% bovine serum albumin and 0.3% Triton TM X-100 for 1 h at room temperature. Sections were incubated overnight at 4°C with the primary antibodies diluted in PBS. Primary antibodies included: rabbit anti-fibronectin, 1:200 (Millipore ab2033); mouse anti-EIIIA-fibronectin, 1:200 (Abcam ab6328); chicken anti-GFAP, 1:500 (Abcam ab4674). For double labeling immunostaining, primary antibodies were incubated sequentially. For staining with the mouse anti-EIIIA-fibronectin (IST9), we used a mouse on mouse (M.O.M.) kit from Vector Laboratories. After staining with primary antibodies, sections were washed with PBS and incubated with Alexa Fluor (488 or 647, 594)-conjugated secondary antibodies (1:400) at room temperature for 2 h, followed by 4′,6-diamidino-2-phenylindole (DAPI) to visualize cell nuclei. Sections were imaged using a Zeiss Axio Observer fluorescence microscope or a laser-scanned confocal microscope (LSM 710, Carl Zeiss, Jena, Germany).