OCLN Antibody / Occludin (FY12345)

Immunohistochemical staining of OCLN using anti-OCLN antibody. 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 ug/ml rabbit anti-OCLN antibody overnight at 4oC. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37oC. The tissue section was developed using an HRP secondary and DAB substrate.

Western blot analysis of OCLN using anti-OCLN antibody. 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-OCLN antibody at 0.5 ug/ml overnight at 4oC, 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 was developed using enhanced chemiluminescent. The expected molecular weight of OCLN is ~59 kDa.

Immunoprecipitation of OCLN in HepG2 whole cell lysate. Western blot analysis of OCLN using anti-OCLN antibody. Lane 1: HepG2 whole cell lysates (30ug), Lane 2: Rabbit control IgG instead of anti-OCLN antibody in HepG2 whole cell lysate, Lane 3: anti-OCLN antibody (2ug) + HepG2 whole cell lysate (500ug). After electrophoresis, proteins were transferred to a membrane. Then the membrane was incubated with rabbit anti-OCLN antibody at a dilution of 0.5 ug/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. The expected molecular weight of OCLN is ~59 kDa.

Immunohistochemical staining of OCLN using anti-OCLN antibody. 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 ug/ml rabbit anti-OCLN antibody overnight at 4oC. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37oC. The tissue section was developed using an HRP secondary and DAB substrate.

Immunohistochemical staining of OCLN using anti-OCLN antibody. 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 ug/ml rabbit anti-OCLN antibody overnight at 4oC. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37oC. The tissue section was developed using an HRP secondary and DAB substrate.