ZO-1 Polyclonal Antibody detects ZO-1 protein by western blot analysis. A. 30 µg 293T whole cell lysate/extract. B. 30 µg A431 whole cell lysate/extract. C. 30 µg HeLa whole cell lysate/extract.5 % SDS-PAGE. ZO-1 Polyclonal Antibody (Product # PA5-21958) dilution: 1:500.

Western Blot using ZO-1 Polyclonal Antibody (Product # PA5-21958). Sample (30 µg of whole cell lysate). A: HCT116.5% SDS PAGE. ZO-1 Polyclonal Antibody (Product # PA5-21958) diluted at 1:1,000.

Immunofluorescence analysis of ZO-1 was performed using 90% confluent log phase Caco-2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with ZO-1 Polyclonal Antibody (Product # PA5-21958) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing Junctional localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.