Rabbit anti mouse cd11b antibody

After blood samples were collected, all rats were transcardially perfused with 150-200 mL saline, followed by 200 mL phosphate buffered saline (PBS, pH = 7.4) for 20-30 min. Liver and brain specimens were removed and fixed in 10% neutral buffered formaldehyde for 24 hours, and embedded in paraffin. Specimens were then cut to a thickness of 5 μm using a vibratome. The hematoxylin-eosin (H&E) staining of liver samples was done by the conventional method 22 (link). For brain immunohistochemistry, free-floating immunohistochemistry was performed as described by previous studies 2 (link), 28 (link). For staining of microglia, rabbit anti-mouse CD11b antibody (1:500; Abcam, Cambridge, UK) was used as the primary antibody. The secondary antibody was a goat anti-rabbit immunoglobulin G (1:500; Vector, Burlingame, California, USA), which was detected by using the horseradish peroxidase (HRP) method. The detailed method can be found in supplementary materials. As previously described 1 (link), 2 (link), 5 , 21 (link), the typical morphology of resting microglia is ramified, while the activated microglia is transferred into a rod or ameboid shape. CD11b immunoreactive microglia cells in basal ganglia were quantified according to Rodrigo et al's method 2 (link). Histomorphometric analysis was performed with a photomicroscope and digital camera (Olympus IX71, Tokyo, Japan).