Goat polyclonal antibody against iba 1

Each brain was cut into 4‐μm‐thick sections for immunohistochemical studies after MI. The slices were incubated with primary antibodies: goat polyclonal antibody against Iba‐1(1:200, Abcam, Cambridge, UK) and anti‐CD34 rabbit polyclonal antibody (1:2500, Abcam) at 4°C overnight, after which DAB substrate (Vector Laboratories, Burlingame, CA, USA) and haematoxylin were used in order for staining.

To detect BBB leakage in 3-NP-injected rats, experimental rats (n = 3 animals) were deeply anesthetized on days 3 and 7 after 3-NP injection and intravenously administered fluorescein isothiocyanate (FITC)-albumin (2 mg diluted in 0.1 ml saline; Sigma-Aldrich) via the tail vein. One hour after injection, animals were deeply anesthetized and perfused transcardially with fixative, as described above. Coronal cryostat sections (25-μm-thick) from animals injected with FITC-albumin were incubated overnight at 4°C with a mixture of rabbit polyclonal antibody against GRP78 (1:2000; Abcam) and one of following antibodies: goat polyclonal antibody against Iba1 (1:500; Abcam), mouse monoclonal antibody against RECA1 (1:200; Bio-Rad), or mouse monoclonal antibodies against GFAP (1:700; Millipore). This was followed by 2-h incubation with appropriate secondary antibodies as follows: Cy3-conjugated donkey anti-goat antibody (1:2000; Jackson ImmunoResearch), Cy3-conjugated goat anti-mouse antibody (1:2000; Jackson ImmunoResearch), or Alexa Fluor 647-tagged donkey anti-rabbit antibody (1:300; Thermo Fisher). Counterstaining of cell nuclei was carried out using DAPI for 10 min.