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Immunofluorescent analysis of Occludin was performed on 90% confluent log phase Caco-2 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with ABfinity™ Occludin recombinant rabbit oligoclonal antibody (Product # 710192) at a dilution of 1:500 in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Flour® 488 goat anti-rabbit IgG secondary antibody (Product # A11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c is a merged image showing cell junction localization and panel d is a control without primary antibody. The images were captured using a Nikon microscope at 20X magnification.
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Peptide corresponding to amino acids 387-403 of human occuludin|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:100|
|Western Blot (WB)||1:500-1:5000|
* 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.
This antibody is predicted to react with equine, mouse, rabbit and rat based on sequence homology.
ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale and purified with Protein A.
ABfinity™ oligoclonal antibodies comprise a selection of multiple different recombinant monoclonal antibodies, providing the best of both worlds—the sensitivity of a polyclonal antibody with the specificity of a monoclonal, all delivered with the consistency only found in a recombinant antibody. While functionally the same as a polyclonal antibody—recognizing multiple epitope sites on the target and producing higher detection sensitivity for low abundance targets when compared with monoclonal antibodies—an oligoclonal antibody has a known mixture of light and heavy chains. This exact population can be produced in every lot, circumventing the biological variability typically associated with polyclonal antibody production.
Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.
Tight junction group of proteins are a cell-to-cell adhesion structure in epithelial cells that constitute the epithelial junctional complex with adherens junctions and desmosomes. Tight junction strands are mainly composed of claudins, occludin, and JAM. Occludin is a 65 kDa protein and is considered to be vital in the assembly and maintenance of tight junctions. At the tight junction, occludin associates with membrane peripheral protein ZO-1. The expression of occludin varies greatly on the type of tissue. It is continuously and highly expressed in brain whereas non-neural tissues show lower expression and discontinuous distribution. Overall structural features of the occludin protein are highly conserved in all the species examined. Under-expression of tight junction proteins, including occludin, is a key molecular abnormality responsible for the increased permeability of tumor endothelial tight junctions, which contributes to brain tumor edemas
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
occludin; OCLN; phosphatase 1, regulatory subunit 115; tight junction protein occludin
AI503564; BLCPMG; Ocl; OCLN; PPP1R115