Overview

  • Product name
  • 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
  • Immunogen

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

    (Peptide available as ab34440.)

  • Positive control
    • WB: HAP1 and HepG2 Cell lysates

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
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

  • 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

  • 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 kDa



    Lanes 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.

  • 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:
  • Wu D  et al. Formononetin Administration Ameliorates Dextran Sulfate Sodium-Induced Acute Colitis by Inhibiting NLRP3 Inflammasome Signaling Pathway. Mediators Inflamm 2018:3048532 (2018). Read more (PubMed: 29507526) »
  • Guo W  et al. Analysis of differential expression of tight junction proteins in cultured oral epithelial cells altered by Porphyromonas gingivalis, Porphyromonas gingivalis lipopolysaccharide, and extracellular adenosine triphosphate. Int J Oral Sci 10:e8 (2018). Read more (PubMed: 29319048) »

See all 66 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.

...

Read More
Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Dog Cell (MDCK (epithelial))
Permeabilization
Yes - 0.1% TritonX
Specification
MDCK (epithelial)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Victoria Wu

Verified customer

Submitted Feb 26 2018

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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1-10 of 21 Abreviews or Q&A

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