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    occludin-antibody-ab31721.pdf

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Signal Transduction Cytoskeleton / ECM Cell Adhesion Tight Junctions
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Anti-Occludin antibody (ab31721)

  • Datasheet
  • SDS
Reviews (16)Q&A (10)References (120)

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Western blot - Anti-Occludin antibody (ab31721)
  • Western blot - Anti-Occludin antibody (ab31721)

Key features and details

  • Rabbit polyclonal to Occludin
  • Suitable for: WB
  • Reacts with: Human, Recombinant fragment
  • Isotype: IgG

You may also be interested in

Protein
Product image
Recombinant Human Occludin protein (ab114189)
Primary
Product image
Anti-Claudin 1 antibody (ab129119)
Secondary
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)

View more associated products

Overview

  • 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 & Species

    Application Species
    WB
    Human
    Recombinant fragment
    See all applications and species data
  • 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.)
  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

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

    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

    • Signal Transduction
    • Cytoskeleton / ECM
    • Cell Adhesion
    • Tight Junctions

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Positive Controls

    • Recombinant Human Occludin protein (ab114189)
  • Recombinant Protein

    • Recombinant Human Occludin protein (ab114189)

Applications

The Abpromise guarantee

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
WB
Human
Recombinant fragment
All applications
Dog
Pig
Application Abreviews Notes
WB (4)
1/250. Detects a band of approximately 62 kDa (predicted molecular weight: 59 kDa).
Notes
WB
1/250. Detects a band of approximately 62 kDa (predicted molecular weight: 59 kDa).

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.
  • Target information above from: UniProt accession Q16625 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 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
    • 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

    • Western blot - Anti-Occludin antibody (ab31721)
      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 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.

    • Western blot - Anti-Occludin antibody (ab31721)
      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

    Protocols

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (120)

    Publishing research using ab31721? Please let us know so that we can cite the reference in this datasheet.

    ab31721 has been referenced in 120 publications.

    • Wang D  et al. Soluble CD146, a cerebrospinal fluid marker for neuroinflammation, promotes blood-brain barrier dysfunction. Theranostics 10:231-246 (2020). PubMed: 31903117
    • Guo C  et al. Bilobalide reversibly modulates blood-brain barrier permeability through promoting adenosine A1 receptor-mediated phosphorylation of actin-binding proteins. Biochem Biophys Res Commun 526:1077-1084 (2020). PubMed: 32312522
    • Dong W  et al. Mesenchymal-endothelial transition-derived cells as a potential new regulatory target for cardiac hypertrophy. Sci Rep 10:6652 (2020). PubMed: 32313043
    • Cohen J  et al. The Wave complex controls epidermal morphogenesis and proliferation by suppressing Wnt-Sox9 signaling. J Cell Biol 218:1390-1406 (2019). PubMed: 30867227
    • Wu H  et al. Mumps virus infection disrupts blood-testis barrier through the induction of TNF-a in Sertoli cells. FASEB J 33:12528-12540 (2019). PubMed: 31450968
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 26 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Occludin antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 07 2012

    Question

    Thanks so much for your cooperation in this matter.

    Read More

    Abcam community

    Verified customer

    Asked on Jul 10 2012

    Answer

    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.

    If there is anything else I can help you with, please let me know.

    Read More

    Abcam Scientific Support

    Answered on Jul 10 2012

    Western blot abreview for Anti-Occludin antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Pig Tissue lysate - other (Jejunum mucosal scraping)
    Gel Running Conditions
    Reduced Denaturing (10% gel)
    Loading amount
    25 µg
    Specification
    Jejunum mucosal scraping
    Blocking step
    Licor intercept blocking buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1µg/mL · Temperature: 27°C
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 26 2021

    Western blot abreview for Anti-Occludin antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Human coronary artery endothelial cells, HeLa and)
    Gel Running Conditions
    Reduced Denaturing (4-20% gradient)
    Loading amount
    50 µg
    Specification
    Human coronary artery endothelial cells, HeLa and
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More

    Sylwia Kabacik

    Verified customer

    Submitted Dec 17 2020

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Occludin antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Keratinocyte)
    Permeabilization
    Yes - PBS-triton 0.5%
    Specification
    Keratinocyte
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
    Fixative
    Acetone
    Read More

    Abcam user community

    Verified customer

    Submitted Jul 15 2020

    Western blot abreview for Anti-Occludin antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Dog Cell lysate - whole cell (MDCK)
    Gel Running Conditions
    Reduced Denaturing (4-12% Bis Acrylamide)
    Loading amount
    45 µg
    Specification
    MDCK
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More

    Dr. Tony Durand

    Verified customer

    Submitted Jul 10 2020

    IHC - Wholemount abreview for Anti-Occludin antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    IHC - Wholemount
    Sample
    Pig Tissue (jejunum tissue from a Landrace pig)
    Specification
    jejunum tissue from a Landrace pig
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 14 2019

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Occludin antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Victoria Wu

    Verified customer

    Submitted Feb 26 2018

    Immunohistochemistry (Frozen sections) abreview for Anti-Occludin antibody

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Dec 08 2015

    ELISA abreview for Anti-Occludin antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    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)
    Read More

    Abcam user community

    Verified customer

    Submitted Jun 03 2015

    1-10 of 26 Abreviews or Q&A

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