Rabbit anti ifn beta receptor

The proteins of ipsilateral/right hemispheres were extracted by cytoplasmic extraction reagents (Pierce Biotechnology, Rockford, IL, USA). Proteins were then loaded (50 μg) and separated by SDS-PAGE gel electrophoresis. After blocking with 5% nonfat milk for 1.5 h, the membranes were incubated overnight at 4 °C with the primary antibodies: rabbit anti-DRD1 (1:1000, Abcam), goat anti-IFN-beta (1:300, Santa Cruz), rabbit anti-IFN-beta receptor (1:500, Santa Cruz), rabbit anti-STAT1(1:2000, Cell Signaling), goat anti-p-STAT1 (1:500, Santa Cruz), rabbit anti-NLRP3 (1:1000, MyBioSource), rabbit anti-caspase1 (1:1000, NOVUS Biologicals), rabbit anti-IL-1beta (1:2000, Cell Signaling), and rabbit anti-β-actin (1:2000, Santa Cruz). β-actin served as the loading control. The membranes were processed with the respective horseradish peroxidase-labeled secondary antibody (Santa Cruz Biotechnology). Bands were visualized using the ECL detection reagents (Amersham Biosciences). The relative density of protein was analyzed by ImageJ software.

Immunofluorescence staining was performed according to previous study [17 (link)]. A series of coronal brain sections (10 μm thick) were blocked in 5% bovine serum albumin for 2 h at room temperature and then were incubated with rabbit anti-DRD1 (1:200, Abcam), rabbit anti-IFN-beta receptor (1:50, Santa Cruz), goat anti-ionized calcium-binding adapter molecule 1 (Iba 1, 1:200, Abcam), and rabbit anti-MPO (1:100, Abcam) at 4 °C overnight. After being washed three times with PBS, the sections were incubated with appropriate fluorescence-conjugated secondary antibodies (1:300, Jackson ImmunoResearch) for 2 h at room temperature. The sections were rinsed three times for 5 min each with PBS. The loci around hematoma (black triangle, Fig. 4a) of stained sections were examined with a fluorescence microscope (Leica DMi8). Microphotographs were analyzed with LASX software.