Human Occludin Antibody

Occludin in HUVEC Human Cells. Occludin was detected in immersion fixed HUVEC human umbilical vein endothelial cells using Mouse Anti-Human Occludin Monoclonal Antibody (Catalog # MAB7074) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the Northern-Lights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Human Occludin by Western Blot Analysis of barrier formation in endothelial monolayer.(A) Expression of endothelial junctional molecules CD31, CD144 and ZO-1 (from left to right) and actin (far right) in hCMVEC monolayer 48 h after seeding. (B) The permeability of hCMVEC monolayer to FITC-dextran 48 h after seeding (mean ± SEM, n = 4). (C) Time course analysis of endothelial tight junction molecule occludin expression at 24, 48 and 72 h post seeding of hCMVECs using Western blot using a mouse monoclonal anti-occludin (left) and rabbit polyclonal anti-occludin (right). (D) Real time measurement of electrical resistance across the endothelial monolayer as an indicator of endothelial barrier integrity. The barrier resistance (Rb) and the basolateral adhesion (Alpha) due to focal adhesions can be modelled using the ECIS Z theta software when the experiment is run using multi-frequency mode (250 to 64000 Hz). The modelling reveals that the basolateral adhesion occurs very fast and junction formation begins around 10 hours after seeding. Overall barrier resistance is the summation of both Rb and alpha, but this software function can determine whether barrier changes are predominantly due to the Rb. Data are mean ± SEM (n = 4). Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0180267), licensed under a CC-BY license. Not internally tested by R&D Systems.