2D HIO were fixed in 4% paraformaldehyde and incubated with the following primary antibodies: mouse anti-CFTR, clone 13-1 (R & D Systems), 1:200 dilution; rabbit anti-zonula occludens-1 (ZO1; Invitrogen), 1:300 dilution in 5% of BSA-PBS; polyclonal goat anti-Ezrin (Santa Cruz), 1: 100 dilution, and then secondary antibodies: Alexa 488 or 594 HRP conjugated to anti-mouse IgG, anti-rabbit IgG (Invitrogen); diluted 1:400 in 5% BSA-PBS. DAPI (4′,6-diamidino-2-phenylindole; Invitrogen) was used to stain the nuclei. Samples were imaged with Olympus spinning disc confocal microscope (Du et al, 2015 (link)).
Rabbit anti zonula occludens 1 zo1
Manufactured by Thermo Fisher Scientific
Rabbit anti-zonula occludens-1 (ZO1) is a primary antibody that recognizes the zonula occludens-1 (ZO1) protein. ZO1 is a tight junction-associated protein involved in the regulation of cell-cell adhesion and permeability. This antibody is commonly used in immunostaining and western blotting applications to detect and analyze the expression and localization of ZO1.
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Lab products found in correlation
Dapi 4 6 diamidino 2 phenylindole, by Thermo Fisher Scientific (1 mentions)
Spinning disc confocal microscope, by Olympus (1 mentions)
Rhodamine phalloidin, by Thermo Fisher Scientific (1 mentions)
Rabbit anti occludin, by Thermo Fisher Scientific (1 mentions)
Mouse anti claudin 5, by Thermo Fisher Scientific (1 mentions)
2 protocols using rabbit anti zonula occludens 1 zo1
1
Immunofluorescent Imaging of 2D HIO
2
Immunofluorescence Analysis of Tight Junctions
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HBMECs were fixed in 4% formaldehyde for 20 min at room temperature and permeabilized in 0.1% Triton X-100 in PBS for 15 min at 4 °C. Cells were incubated with antibodies such as rabbit anti-occludin (1:100, Invitrogen), mouse anti-claudin-5 (1:50, Invitrogen), or rabbit anti-zonula occludens-1 (ZO-1, 1:200, Invitrogen) overnight at 4 °C. The cells were incubated with secondary antibodies conjugated with Alexa Fluor for 1 h at room temperature. Actin filaments were monitored with rhodamine phalloidin (Molecular Probes) for 30 min. Cells were mounted using Dako mounting reagent and were observed using a fluorescence microscope (Zeiss; ×400).
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