Immunofluorescence analysis of Occludin Antibody, Alexa Fluor® 594 conjugate (OC-3F10) 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, Alexa Fluor® 594 conjugate (OC-3F10)(331594) at 1µg/mL in 1% BSA and incubated for 3 hours at room temperature (Panel a: red). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). Panel c: red). Panel c is a merged image showing cell junctional localization. Panel d is a no primary antibody control. The images were captured at 40X magnification.
|Tested species reactivity||Dog, Human, Mouse, Rat|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||GST fusion protein consisting of the C-terminal region (~150aa) of human occludin.|
|Conjugate||Alexa Fluor® 594|
|Storage buffer||PBS, pH 7.4, with 4mg/ml BSA|
|Contains||0.1% sodium azide|
|Storage Conditions||4° C, store in dark|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|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 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.
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