Western blot analysis of Occludin was performed by loading 10 ug of CaCO2 lysate per well onto a 4-12% Bis-Tris polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with an Occludin purified monoclonal antibody (Product # 331500) at a dilution of 1 ug/ml, 0.5 ug/ml, and 0.25 ug/ml for 1 hour at room temperature on a rocking platform and washed in TBS-0.1% Tween-20. Goat Anti-Mouse IGG (H+L) HRP Conjugated secondary antibody (Product# 31430) was used at a concentration of 1:20,000 and incubated for 30 minutes on a rocking platform. Super Signal West Dura substrate (Product #34076) was incubated for 5 minutes on a rocking platform. Detection was performed using the GBox. The 5th isoform of Occludin was detected at ~23 kDa.
|Tested species reactivity||Dog, Human, Mouse, Rat|
|Published species reactivity||Dog, Rat, Human|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||GST fusion protein consisting of the C-terminal region (~150aa) of human occludin.|
|Storage buffer||PBS, pH 7.4, with 4mg/ml BSA, 40% glycerol|
|Contains||0.1% Proclin 300|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|ELISA (ELISA)||0.1-1.0 ug/ml|
|Western Blot (WB)||0.1-1.0 ug/ml|
* 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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
12/15-Lipoxygenase-dependent ROS production is required for diet-induced endothelial barrier dysfunction.
33-1520 was used in immunocytochemistry and western blot to examine the mechanisms of 15(S)-HETE-induced endothelial cell barrier dysfunction
|Chattopadhyay R,Tinnikov A,Dyukova E,Singh NK,Kotla S,Mobley JA,Rao GN||Journal of lipid research (56:562)||2015|
HIV-1 Tat C modulates expression of miRNA-101 to suppress VE-cadherin in human brain microvascular endothelial cells.
33-1520 was used in western blot to demonstrate the effect of HIV-1 Tat C on miRNA-101 expression in human brain microvascular endothelial cells
|Mishra R,Singh SK||The Journal of neuroscience : the official journal of the Society for Neuroscience (33:5992)||2013|
|Human||Not Cited||Differential effects of flavonoids on barrier integrity in human intestinal Caco-2 cells.||Noda S,Tanabe S,Suzuki T||Journal of agricultural and food chemistry (60:4628)||2012|
|Rat||Not Cited||Role of megalin (gp330) in transcytosis of thyroglobulin by thyroid cells. A novel function in the control of thyroid hormone release.||Marinò M,Zheng G,Chiovato L,Pinchera A,Brown D,Andrews D,McCluskey RT||The Journal of biological chemistry (275:7125)||2000|
|Protein kinase C¿ phosphorylates occludin and promotes assembly of epithelial tight junctions.||Jain S,Suzuki T,Seth A,Samak G,Rao R||The Biochemical journal (437:289)||2011|