Anti-ZO1 tight junction protein antibody (ab59720)

Overview

  • Product name
    Anti-ZO1 tight junction protein antibody
    See all ZO1 tight junction protein primary antibodies
  • Description
    Rabbit polyclonal to ZO1 tight junction protein
  • Specificity
    We are re-evaluating this product to ensure it meets our strict quality criteria. Unfortunately it will not be available to order during this time. We recommend ab216880 as an alternative rabbit polyclonal which works in ICC/IF, WB and IHC-P.
  • Tested applications
    Suitable for: ICC/IF, Flow Cyt, WBmore details
  • Species reactivity
    Reacts with: Chicken, Dog, Human
    Predicted to work with: Mouse, Pig
  • Immunogen

    Corresponding to amino acids 432-1150 of Human ZO1 tight junction protein

  • General notes

    ab59720 recognises both ZO1 tight junction alpha-minus and alpha-plus isoforms.
    The protein is abundant in endothelial cells and the highly specialized epithelial junctions.

    Customers report variable results in IHC-Fr and IHC-P application; though we have nice image for IHC-Fr. If you would like to use ab59720 in IHC-Fr then we suggest using ethanol for fixation instead of acetone.

    IHC-Fr and IHC-P are therefore not covered by Abpromise guarantee.

    The immunogen sequence for this product has 92.5% homology with the mouse sequence however in our hands we have not been able to produce positive data with mouse samples.

    We have also received mixed feedback for Pig cross reactivity so we no longer guarantee this species. One of the reviewer have successfully used this antibody with pig samples so we are keen to receive more feedback from other customers.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer
    Preservative: 0.02% Sodium Azide
    Constituents: 0.1% BSA, PBS
  • Concentration information loading...
  • Purification notes
    0.2µm filtered.
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab59720 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
ICC/IF Use a concentration of 1 µg/ml.

Fix cells with 4% paraformaldehyde and permeabilize with PBS + 0.1% Saponin (+ 0.5% BSA).

 

Flow Cyt 1/50.

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

WB 1/50 - 1/250. Predicted molecular weight: 194 kDa.

Abcam recommends BSA blocking.

Target

  • Function
    The N-terminal may be involved in transducing a signal required for tight junction assembly, while the C-terminal may have specific properties of tight junctions. The alpha domain might be involved in stabilizing junctions.
  • Tissue specificity
    The alpha-containing isoform is found in most epithelial cell junctions. The short isoform is found both in endothelial cells and the highly specialized epithelial junctions of renal glomeruli and Sertoli cells of the seminiferous tubules.
  • Sequence similarities
    Belongs to the MAGUK family.
    Contains 1 guanylate kinase-like domain.
    Contains 3 PDZ (DHR) domains.
    Contains 1 SH3 domain.
    Contains 1 ZU5 domain.
  • Domain
    The second PDZ domain mediates interaction with GJA12.
  • Post-translational
    modifications
    Phosphorylated. Dephosphorylated by PTPRJ.
  • Cellular localization
    Cell membrane. Cell junction > tight junction. Movement of ZO-1 from the cytoplasm to membrane is an early event occurring concurrently with cell-cell contact.
  • Information by UniProt
  • Database links
  • Alternative names
    • Tight junction protein 1 antibody
    • Tight junction protein ZO-1 antibody
    • Tight junction protein ZO1 antibody
    • TJP1 antibody
    • zo-1 antibody
    • Zo1 antibody
    • ZO1_HUMAN antibody
    • Zona occludens 1 antibody
    • Zona occludens 1 protein antibody
    • Zona occludens protein 1 antibody
    • Zonula occludens 1 protein antibody
    • Zonula occludens protein 1 antibody
    see all

Anti-ZO1 tight junction protein antibody images

  • Immunofluorescence staining in Huvec cells.

  • Flow cytometry with Huvec cells. The black line represents the cells only, the red line the control and the blue line ab59720 at 10 μg/ml.

  • ab59720 staining ZO1 in Caco-2 cells treated with (±)-palmitoylcarnitine chloride (ab141122), by ICC/IF. Membranar ZO1 expression loss correlates with increased concentration of (±)-palmitoylcarnitine chloride, as described in literature.
    The cells were incubated at 37°C for 1 hour in media containing different concentrations of ab141122 ((±)-palmitoylcarnitine chloride) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab59720 (0.5 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

  • ICC/IF image of ab59720 stained MCF-7 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 (ab59720, 1µg/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.

  • Immunofluorescence analysis of HUVEC cells, staining ZO1 tight junction protein with ab59720.

    Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 1% BSA for 30 minutes at 25°C. Samples were incubated with primary antibody (1/200 in diluent) for 16 hours at 4°C. An AlexaFluor®488-conjugated goat anti-rabbit polyclonal IgG (1/500) was used as the secondary antibody.

    See Abreview

  • Immunofluorescence analysis of murine peripheral retinal pigmented epithelium (RPE) cells, staining ZO1 tight junction protein with ab59720.

    Cells were fixed with 4% paraformaldehyde for 10 minutes at room temperature. Cells were blocked in 10% BSA and 0.5% Triton-X 100 for 2 hours at room temperature before being incubated with primary antibody (1/100) in blocking solution overnight at room temperature. An AlexaFluor®-conjugated anti-rabbit IgG was used as the secondary antibody.

References for Anti-ZO1 tight junction protein antibody (ab59720)

This product has been referenced in:
  • Li J  et al. Nicotinic acid inhibits glioma invasion by facilitating Snail1 degradation. Sci Rep 7:43173 (2017). Read more (PubMed: 28256591) »
  • Chen YC  et al. High mobility group box 1-induced epithelial mesenchymal transition in human airway epithelial cells. Sci Rep 6:18815 (2016). WB, ICC/IF ; Human . Read more (PubMed: 26739898) »

See all 40 Publications for this product

Product Wall

Application
IHC - Wholemount
Sample
Mouse Tissue (intestine organoids)
Specification
intestine organoids
Username

Abcam user community

Verified customer

Submitted Mar 29 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Quail Tissue sections (testis)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer
Permeabilization
No
Specification
testis
Blocking step
Milk as blocking agent for 5 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Formaldehyde
Username

Mrs. Ilknur Tekdemir

Verified customer

Submitted Feb 20 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
IHC - Wholemount
Sample
Mouse Embryo (lung)
Specification
lung
Username

Abcam user community

Verified customer

Submitted Dec 08 2016

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Heart)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Buffer
Permeabilization
Yes - 0.2% Triton X-100
Specification
Heart
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Oct 26 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (brain)
Permeabilization
Yes - 0.25% tx-100
Specification
brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Feb 23 2016

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Embryonic mouse brain tissue)
Permeabilization
Yes - triton X
Specification
Embryonic mouse brain tissue
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Dr. Bhavin Shah

Verified customer

Submitted Oct 19 2015

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate
Permeabilization
Yes - 0.1% Triton
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Mar 13 2015

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Sheep Tissue sections (1-2 month old testis)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 0.01M citric acide/sodium citrate
Permeabilization
No
Specification
1-2 month old testis
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
4%PFA
Username

Abcam user community

Verified customer

Submitted Jan 14 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (MCF7 cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
MCF7 cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jan 07 2015

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Zebrafish Tissue sections (Retina, Outer limiting membrane)
Permeabilization
Yes - triton X
Specification
Retina, Outer limiting membrane
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Sep 05 2014

1-10 of 32 Abreviews or Q&A

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