概述

  • 产品名称Anti-Occludin抗体
    参阅全部 Occludin 一抗
  • 描述
    兔多克隆抗体to Occludin
  • 特异性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.
  • 经测试应用适用于: IHC-FoFr, WB, In-Cell ELISA, ICC/IF, IHC-Pmore details
  • 种属反应性
    与反应: Human, Pig
    预测可用于: Dog
  • 免疫原

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

    (Peptide available as ab34440.)

  • 阳性对照
    • ab31721 gave a positive signal in Colon (Human), Liver (Human), and Kidney (Human) Tissue Lysates.

性能

  • 形式Liquid
  • 存放说明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.
  • 存储溶液Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • 纯度Immunogen affinity purified
  • 克隆多克隆
  • 同种型IgG
  • 研究领域

应用

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.

应用 Ab评论 说明
IHC-FoFr Use at an assay dependent concentration. PubMed: 20978075
WB Use a concentration of 1 µg/ml. Detects a band of approximately 63 kDa (predicted molecular weight: 59 kDa).

Abcam recommends using 3% milk as the blocking agent.

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.

靶标

  • 功能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.
  • 组织特异性Localized at tight junctions of both epithelial and endothelial cells. Highly expressed in kidney. Not detected in testis.
  • 疾病相关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.
  • 序列相似性Belongs to the ELL/occludin family.
    Contains 1 MARVEL domain.
  • 结构域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.
  • 翻译后修饰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.
  • 细胞定位Membrane. Cell junction > tight junction.
  • Information by UniProt
  • 数据库链接
  • 别名
    • BLCPMG antibody
    • FLJ08163 antibody
    • FLJ18079 antibody
    • FLJ77961 antibody
    • FLJ94056 antibody
    • MGC34277 antibody
    • Occludin antibody
    • Ocln antibody
    • OCLN_HUMAN antibody
    • Tight junction protein occludin antibody
    see all

Anti-Occludin antibody 图像

  • 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

  • All lanes : Anti-Occludin antibody (ab31721) at 1 µg/ml

    Lane 1 : Rat colon tissue lysate
    Lane 2 : Human colon tissue lysate
    Lane 3 : Human kidney tissue lysate
    Lane 4 : Human skeletal muscle tissue lysate

    Lysates/proteins at 20 µg per lane.


    Performed under reducing conditions.

    Predicted band size : 59 kDa
    Observed band size : 62 kDa (why is the actual band size different from the predicted?)

    Tissue lysates were denatured for 10-15 minutes at 90ºC. ab31721 was incubated overnight at 4ºC and the secondary antibody for 1 hour at RT. Mouse GAPDH loading control was used (red channel) at 1/5000 dilution. Blocking was performed using Licor blocking buffer. 

     

    The immunogen sequence for ab31721 shares 78% homology with the Rat Occludin protein, which is not detected in WB (lane 1). We therefore cannot guarantee reactivity with this species.

  • 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

  • All lanes : Anti-Occludin antibody (ab31721) at 1 µg/ml

    Lane 1 : Human colon tissue lysate - total protein (ab30051)
    Lane 2 : Human liver tissue lysate - total protein (ab29889)
    Lane 3 : Human skeletal muscle tissue lysate - total protein (ab29330)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 59 kDa
    Observed band size : 63 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 240 kDa,32 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 12 minutes

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab31721 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

  • 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.
  • Anti-Occludin antibody (ab31721) at 1 µg/ml (blocked with 3% milk) + Human kidney tissue lysate - total protein (ab30203) at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 59 kDa
    Observed band size : 63 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 26 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 2 minutes

Anti-Occludin antibody (ab31721)参考文献

This product has been referenced in:
  • Cao S  et al. Hydrogen sulfide attenuates brain edema in early brain injury after subarachnoid hemorrhage in rats: Possible involvement of MMP-9 induced blood-brain barrier disruption and AQP4 expression. Neurosci Lett 621:88-97 (2016). WB ; Rat . Read more (PubMed: 27080433) »
  • Zhang C  et al. Catalpol downregulates vascular endothelial-cadherin expression and induces vascular hyperpermeability. Mol Med Rep 13:373-8 (2016). IF . Read more (PubMed: 26549479) »

See all 25 Publications for this product

Product Wall

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

提交于 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
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

提交于 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

提交于 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

提交于 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

提交于 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

提交于 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

提交于 Sep 23 2013

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