Rabbit anti aqp4 antibody

V79 cells expressing M23-AQP4 were cultured on round glass coverslips in 24-well plate (Corning, Inc.) for 24 h. After washing three times with PBS, the cells were blocked with 3% BSA (Merck KGaA) at room temperature for 1 h. Geraldol or DMSO was added in 3% BSA and incubated at room temperature for 15 min, followed by the addition of NMO-IgG (20 µg/ml) or control serum from patients without NMO (1:200) at room temperature for 1 h. V79-AQP4 cells were washed with PBS three times and incubated with Alexa Fluor 555-conjugated goat anti-human IgG secondary antibody (1:400; Invitrogen; Thermo Fisher Scientific, Inc; cat. no. A-21433). For AQP4 immunofluorescence staining, V79-AQP4 cells were fixed in 4% PFA at room temperature for 10 min and permeabilized with 0.5% Triton X-100 at room temperature for 10 min. After blocking with 3% BSA at room temperature for 1 h, the cells were incubated with rabbit anti-AQP4 antibody (1:200; Santa Cruz Biotechnology; cat. no. sc-32739) at room temperature for 1 h. Alexa Fluor 488-conjugated goat anti-rabbit IgG (1:200; Invitrogen; Thermo Fisher Scientific, Inc.; cat. no. A32731) was used as the secondary antibody. Fluorescence images were obtained and observed using fluorescence microscopy (IX71; Olympus Corporation).