Immunofluorescence analysis of occludin Antibody was done on 90% confluent log phase CACO2 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with Occludin Antibody (406100) at 1µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa flour 488 Goat Anti-Rabbit IgG Secondary Antibody (A11008) at a dilution of 1:400 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (A12381). Panel d is a merged image showing cell junctional localization. Panel e is a no primary antibody control. The images were captured at 40X magnification.
|Tested species reactivity||Human|
|Published species reactivity||Rat, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide derived from the N-terminal region of the human, rat, mouse, dog, and orangutan occludin proteins|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:100|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Occludin is a 65 kDa protein that can exist in a variety of phosphorylated forms, ranging up to approximately 82 kDa. This phosphorylation is thought to be involved in regulating both the localization and the function of occludin. Polyunsaturated fatty acids are known to up-regulate occludin expression, increasing the transendothelial cell resistance and reducing the cellular permeability to large molecules. The level of occludin varies greatly depending on tissue; in brain tissue, occludin is highly and continuously expressed at cell-cell contact sites, whereas non-neural tissues show lower expression and discontinuous distribution.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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