Anti-Occludin antibody (ab31721)
- Datasheet
- References (105)
- Protocols
Overview
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Product name
Anti-Occludin antibody
See all Occludin primary antibodies -
Description
Rabbit polyclonal to Occludin -
Host species
Rabbit -
Specificity
New batches of this antibody are no longer guaranteed in ICC/IF, IHC-P or IHC-Fr as they have not passed our stringent batch testing criteria. Please contact customer support for any specific queries. We recommend ab216327 as an alternative for ICC/IF and IHC-P. The immunogen used to raise this antibody has 78% homology with the Mouse Occludin protein and the Rat Occludin protein. Some customers have successfully used ab31721 with Mouse/Rat lysates/tissue, however we have not been successful detecting Occludin in these species in our own testing. -
Tested applications
Suitable for: WB, In-Cell ELISAmore details -
Species reactivity
Reacts with: Human, Pig
Predicted to work with: Dog
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 350 - 450 of Human Occludin.
Read Abcam's proprietary immunogen policy (Peptide available as ab34440.) -
Positive control
- WB: Recombinant Human Occludin protein (ab114189), HAP1 and HepG2 Cell lysates
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab31721 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB | 1/250. Detects a band of approximately 62 kDa (predicted molecular weight: 59 kDa). | |
| In-Cell ELISA | Use at an assay dependent concentration. PubMed: 22689949 |
Target
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Function
May play a role in the formation and regulation of the tight junction (TJ) paracellular permeability barrier. It is able to induce adhesion when expressed in cells lacking tight junctions. -
Tissue specificity
Localized at tight junctions of both epithelial and endothelial cells. Highly expressed in kidney. Not detected in testis. -
Involvement in disease
Defects in OCLN are the cause of band-like calcification with simplified gyration and polymicrogyria (BLCPMG) [MIM:251290]; also known as pseudo-TORCH syndrome. BLCPMG is a neurologic disorder with characteristic clinical and neuroradiologic features that mimic intrauterine TORCH infection in the absence of evidence of infection. Affected individuals have congenital microcephaly, intracranial calcifications, and severe developmental delay. -
Sequence similarities
Belongs to the ELL/occludin family.
Contains 1 MARVEL domain. -
Domain
The C-terminal is cytoplasmic and is important for interaction with ZO-1. Sufficient for the tight junction localization. Involved in the regulation of the permeability barrier function of the tight junction (By similarity). The first extracellular loop participates in an adhesive interaction. -
Post-translational
modificationsPhosphorylated upon DNA damage, probably by ATM or ATR. Dephosphorylated by PTPRJ. The tyrosine phosphorylation on Tyr-398 and Tyr-402 reduces its ability to interact with TJP1. -
Cellular localization
Membrane. Cell junction > tight junction. - Information by UniProt
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Database links
- Entrez Gene: 403844 Dog
- Entrez Gene: 100506658 Human
- Entrez Gene: 397236 Pig
- Omim: 602876 Human
- SwissProt: Q28269 Dog
- SwissProt: Q16625 Human
- Unigene: 592605 Human
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Alternative names
- BLCPMG antibody
- FLJ08163 antibody
- FLJ18079 antibody
see all
Images
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Western blot - Anti-Occludin antibody (ab31721)All lanes : Anti-Occludin antibody (ab31721) at 1/250 dilution
Lane 1 : Wild-type HAP1 whole cell lysate at 40 µg
Lane 2 : OCLN (Occludin) knockout HAP1 whole cell lysate at 40 µg
Lane 3 : HeLa whole cell lysate (Low Occludin expression) at 20 µg
Lane 4 : HepG2 whole cell lysate (High Occludin expression) at 20 µg
Predicted band size: 59 kDaLanes 1 - 4: Merged signal (red and green). Green - ab31721 observed at 59 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab31721 was shown to recognize Occludin in wild-type HAP1 cells as signal was lost at the expected MW in OCLN (Occludin) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and OCLN (Occludin) knockout samples were subjected to SDS-PAGE. Ab31721 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/250 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Western blot - Anti-Occludin antibody (ab31721)Lane 1 : Anti-Occludin antibody [EPR8208] (ab167161) at 1/5000 dilution
Lane 2 : Anti-Occludin antibody (ab31721) at 1/1000 dilution
Lane 3 : Anti-Occludin antibody [EPR20992] (ab216327) at 1/5000 dilution
All lanes :Recombinant Human Occludin protein (ab114189)
Lysates/proteins at 0.025 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 59 kDa
Observed band size: 85 kDa why is the actual band size different from the predicted?Blocking and diluting buffer: 5% NFDM/TBST.
Exposure time: 5.5 seconds
References
This product has been referenced in:
- Kasioumi P et al. Hsp70 (HSP70A1A) downregulation enhances the metastatic ability of cancer cells. Int J Oncol 54:821-832 (2019). Read more (PubMed: 30569142) »
- Zhu L et al. The role of LINC00094/miR-224-5p (miR-497-5p)/Endophilin-1 axis in Memantine mediated protective effects on blood-brain barrier in AD microenvironment. J Cell Mol Med 23:3280-3292 (2019). Read more (PubMed: 30801976) »
