Overview

  • Product name
  • Description
    Rabbit polyclonal to Occludin
  • Specificity
    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. Please contact Abcam Scientific Support for more information.
  • Tested applications
    Suitable for: IHC-FoFr, In-Cell ELISA, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Rat, Human, Pig
    Predicted to work with: Dog
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 350 - 450 of Human Occludin.

    (Peptide available as ab34440.)

  • Positive control
    • IHC-P: Normal human kidney tissue. ICC/IF: MCF7 cells.

Properties

  • 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
    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

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
IHC-FoFr Use at an assay dependent concentration. PubMed: 20978075
In-Cell ELISA Use at an assay dependent concentration. PubMed: 22689949
ICC/IF Use a concentration of 1 - 2 µg/ml.
IHC-P 1/100. PubMed: 19828778

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • 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
    modifications
    Phosphorylated 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
  • Database links
  • Alternative names
    • BLCPMG antibody
    • FLJ08163 antibody
    • FLJ18079 antibody
    • FLJ77961 antibody
    • FLJ94056 antibody
    • MGC34277 antibody
    • Occludin antibody
    • Ocln antibody
    • OCLN_HUMAN antibody
    • Phosphatase 1 regulatory subunit 115 antibody
    • PPP1R115 antibody
    • PTORCH1 antibody
    • Tight junction protein occludin antibody
    see all

Images

  • IHC image of Occludin staining in human normal kidney formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab31721, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • ab31721 staining Occludin in Human Breast cancer cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with methanol/acetone (1:1) and blocked with 0.1% milk for 30 minutes at 25°C. Samples were incubated with primary antibody (1/200 in PBS + 0.1% milk) for 16 hours at 4°C. An Alexa Fluor® 594-conjugated Goat anti-rabbit polyclonal (1/1500) was used as the secondary antibody.

    See Abreview

  • ICC/IF image of ab31721 stained Hek293 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab31721, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References

This product has been referenced in:
  • Chelakkot C  et al. Intestinal Epithelial Cell-Specific Deletion of PLD2 Alleviates DSS-Induced Colitis by Regulating Occludin. Sci Rep 7:1573 (2017). WB, IHC-P, ICC/IF ; Mouse . Read more (PubMed: 28484281) »
  • Kou X  et al. Protective Effect of Tang Wang One Decoction on the Retinal Vessels of Diabetic Rats. Evid Based Complement Alternat Med 2017:8635127 (2017). IHC ; Rat . Read more (PubMed: 28367226) »

See all 55 Publications for this product

Customer reviews and Q&As

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Breast cancer)
Specification
Breast cancer
Fixative
Methanol-Acetone
Permeabilization
No
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 0.1% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Aug 07 2012

Thank you for providing that information.

I am sending a new aliquot of this antibody, and I hope that it will resolve the staining issues that you have been having. Your new order number is * and you should have it tomorrow morning.

...

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Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Brain-Arteriovenous malformation)
Permeabilization
Yes - Triton X-100
Specification
Brain-Arteriovenous malformation
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Dec 08 2015

Abreviews
Application
ELISA
Sample
Human Tissue sections (Brain lysate in TBSX)
Specification
Brain lysate in TBSX
Blocking step
BlockAce as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 24°C
Type
Sandwich (Capture)
Username

Abcam user community

Verified customer

Submitted Jun 03 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate
Sample
Mouse Tissue sections (Brain)
Specification
Brain
Permeabilization
Yes - 0.1% Triton
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Mar 24 2015

Application
Western blot
Loading amount
75 µg
Gel Running Conditions
Reduced Denaturing (12.5%)
Sample
Rat Cell lysate - whole cell (Epithelial cells)
Specification
Epithelial cells
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Oct 23 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Antigen retrieval step
Enzymatic
Sample
Mouse Tissue sections (adult testes)
Specification
adult testes
Permeabilization
No
Fixative
4%PFA
Username

Dr. Qing Wen

Verified customer

Submitted May 01 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 45 minute(s) · Concentration: 1.5% · Temperature: 25°C
Antigen retrieval step
Enzymatic
Sample
Rat Tissue sections (GONAD)
Specification
GONAD
Permeabilization
No
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Sep 23 2013

Vielen Dank, dass Sie sich die Zeit genommen haben, unseren Fragebogen auszufüllen.

Es tut mir leid, dass Sie Probleme mit unserem Antikörper hatten.

Ich habe in unserem Lager nachgefragt und muss Ihnen leider mitteilen, das...

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Thank you for contacting us.

I can confirm that different lots of polyclonal antibodies are always different in their detection of target proteins. That is because the immunreaction is different in changing hosts and different immunisation rou...

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