Anti-Occludin/OCLN Antibody Picoband®

Western blot analysis of Occludin/OCLN using anti-Occludin/OCLN antibody (A01246-4).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human HepG2 whole cell lysates,
Lane 2: human 293T whole cell lysates,
Lane 3: human Jurkat whole cell lysates,
Lane 4: rat liver tissue lysates,
Lane 5: mouse liver tissue lysates,
Lane 6: mouse kidney tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Occludin/OCLN antigen affinity purified polyclonal antibody (A01246-4) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Occludin/OCLN at approximately 59 kDa. The expected band size for Occludin/OCLN is at 59 kDa.

IHC analysis of Occludin/OCLN using anti-Occludin/OCLN antibody (A01246-4).
Occludin/OCLN was detected in a paraffin-embedded section of human stomach tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Occludin/OCLN Antibody (A01246-4) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of Occludin/OCLN using anti-Occludin/OCLN antibody (A01246-4).
Occludin/OCLN was detected in a paraffin-embedded section of human stomach tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Occludin/OCLN Antibody (A01246-4) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

Effect of Huanglian-Jiedu decoction on intestinal barrier in APP/PS1 mice. A Representative image of HE staining in the mouse intestine, and representative images of immunohistochemistry of intestinal Occludin and ZO-1 (Scale bar = 50 μm). B Statistical analysis of the relative expression level of Occludin. C Statistical analysis of the relative expression level of ZO-1. D The expressions of ZO-1 and Occludin proteins in the intestine were detected by Western blotting. E Semi-quantitative analysis of the gray value of Occludin. F Semi-quantitative analysis of the gray value of ZO-1. Data are presented as mean ± SD (n = 3). ## P < 0.01 (vs control group), * P < 0.05 and ** P < 0.01 (vs model group)
Index in PubMed under a CC BY license. PMID: 40770806

Effects of MT water extract on regulating macrophage polarization, maintaining intestinal barrier integrity, and alleviating oxidative stress in the colons of DSS-induced UC mice. (A) IHC staining of iNOS, Arg1, occludin, and ZO-1 expressions in mouse colon tissues. Quantitative analysis of the integrated optical density (IOD) of (B) iNOS, (C) Arg1, (D) occludin, and (E) ZO-1 in each group. The levels of (F) IL-6, (G) IL-10, (H) MDA, and (I) GSH were measured and analyzed. Data were presented as means ± SD for three independent trials. One-way ANOVA followed by Tukey's multiple comparison test was performed to compare the differences between groups. *p < 0.05 versus control and #p < 0.05 versus DSS alone group. Arg1, arginase 1; DSS, dextran sulfate sodium; IL, interleukin; iNOS, inducible nitric oxide synthase; MDA, malondialdehyde; MT, Medulla Tetrapanacis.
Index in Food Frontiers under a CC BY license. DOI: 10.1002/fft2.70164

Immunoprecipitating Occludin/OCLN in HepG2 whole cell lysate.
Western blot analysis of Occludin/OCLN using anti-Occludin/OCLN antibody (A01246-4).
Lane 1: HepG2 whole cell lysates (30ug),
Lane 2: Rabbit control IgG instead of anti-Occludin/OCLN antibody in HepG2 whole cell lysate,
Lane 3: anti-Occludin/OCLN antibody (2μg) + HepG2 whole cell lysate (500μg).
After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-Occludin/OCLN antigen affinity purified polyclonal antibody (A01246-4) at a dilution of 0.5 μg/mL and probed with a mouse anti-rabbit IgG-HRP secondary antibody (Light Chain Specific). The signal is developed using ECL Plus Western Blotting Substrate (Catalog # AR1197). A specific band was detected for Occludin/OCLN at approximately 59 kDa. The expected band size for Occludin/OCLN is at 59 kDa.

IHC analysis of Occludin/OCLN using anti-Occludin/OCLN antibody (A01246-4).
Occludin/OCLN was detected in a paraffin-embedded section of human stomach tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Occludin/OCLN Antibody (A01246-4) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.