Protein was isolated in situ, and western analysis performed as previously described [27 (link)]. Blots were probed using antibodies directed to Claudin-1, Occludin-1, ZO-1 (Thermo Fisher Scientific, North Ryde, Sydney, NSW, Australia), LC3, poly (ADP)-ribose polymerase (PARP), Sequestosome (Cell Signaling Technology, Boston, MA), Bcl2, NF-κβ (Santa Cruz Biotechnology, Dallas, TX) and β-actin (Sigma-Aldrich) followed by matching horseradish peroxidase-conjugated secondary antibodies (R&D Systems, MN, USA). Detection was performed using ECL Prime chemiluminescent substrate (GE Healthcare, Buckinghamshire, UK). Densitometry of histogram analyses was performed using Multi Gauge software (V3.1 Fugifilm, Tokyo, Japan). Density scores were normalized to both β-actin and the untreated control, and analyzed using a bootstrapped gamma regression model (log link) and stratified by replicate. The statistical analysis was performed using R statistical software (release 3.2.3) and results expressed as relative abundance.
Occludin 1
Manufactured by Thermo Fisher Scientific
Sourced in Australia, United States
Occludin-1 is a laboratory equipment product designed for use in research applications. It is a key component of tight junctions, which are specialized cell-cell adhesion complexes that play a critical role in the regulation of paracellular permeability. Occludin-1 can be used to study the structure and function of tight junctions in various biological systems.
Automatically generated - may contain errors
Lab products found in correlation
β actin, by Merck Group (2 mentions)
Ecl prime chemiluminescent substrate, by GE Healthcare (1 mentions)
Nf κβ, by Santa Cruz Biotechnology (1 mentions)
Bcl 2, by Santa Cruz Biotechnology (1 mentions)
Horseradish peroxidase conjugated secondary antibodies, by R&D Systems (1 mentions)
Claudin 1, by Thermo Fisher Scientific (1 mentions)
Multi gauge software, by Fujifilm (1 mentions)
2 protocols using occludin 1
1
Western Blot Analysis of Tight Junction Proteins
2
Quantifying Tight Junction Proteins
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Protein was isolated from differentiated AECs within transwells in situ, and Western blot analysis was performed as previously described.16 (link) Blots were probed using antibodies directed to Claudin-1, Occludin-1, ZO-1 (all; Thermo Fisher Scientific, Waltham, MA, USA), and β-actin (Sigma-Aldrich Co., St Louis, MO, USA). Densitometry of histogram analyses was performed using Multi Gauge software (V3.1; Fujifilm, Tokyo, Japan). Density scores were analyzed by a gamma (log link) mixed-model regression to allow for correlated treatment responses within each culture. Results were normalized to both β-actin and the biological control and expressed as relative abundance. The statistical analysis was performed using R statistical software (release 3.2.3) for N=6 cultures.
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