Anti-ZO1 tight junction protein antibody [mAbcam 61357] (ab61357)

Overview

  • Product nameAnti-ZO1 tight junction protein antibody [mAbcam 61357]
    See all ZO1 tight junction protein primary antibodies
  • Description
    Mouse monoclonal [mAbcam 61357] to ZO1 tight junction protein
  • Tested applicationsSuitable for: Flow Cyt, WB, IPmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Dog
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 750 - 850 of Human ZO1 tight junction protein.

    (Peptide available as ab93087.)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: HepG2; HEK293; A431; Caco2.
  • General notes

    Alternative versions available:
    Anti-ZO1 tight junction protein antibody (FITC) [mAbcam 61357] (ab150266)

Properties

  • FormLiquid
  • Storage instructionsShipped 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 bufferPreservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • PurityIgG fraction
  • ClonalityMonoclonal
  • Clone numbermAbcam 61357
  • MyelomaSp2/0-Ag14
  • IsotypeIgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab61357 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
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
WB Use a concentration of 0.1 - 1 µg/ml. Detects a band of approximately 240 kDa (predicted molecular weight: 195 kDa).

Abcam recommends using BSA as the blocking agent.

IP Use a concentration of 5 µg/ml.

Target

  • FunctionThe 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 specificityThe 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 similaritiesBelongs to the MAGUK family.
    Contains 1 guanylate kinase-like domain.
    Contains 3 PDZ (DHR) domains.
    Contains 1 SH3 domain.
    Contains 1 ZU5 domain.
  • DomainThe second PDZ domain mediates interaction with GJA12.
  • Post-translational
    modifications
    Phosphorylated. Dephosphorylated by PTPRJ.
  • Cellular localizationCell 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 [mAbcam 61357] images

  • ZO1 tight junction protein was immunoprecipitated using 0.5mg Hek293 whole cell extract, 5µg of Mouse monoclonal to and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

    The antibody was incubated under agitation with Protein G beads for 10min, Hek293 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab61357.

    Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.

    Band: 240kDa, non specific band - 90kDa: We are unsure as to the identity of this extra band; ZO1 tight junction protein

  • All lanes : Anti-ZO1 tight junction protein antibody [mAbcam 61357] (ab61357) at 5 µg/ml

    Lane 1 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 4 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 195 kDa
    Observed band size : 240 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 38 kDa,44 kDa,48 kDa,85 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 30 seconds
    The 240-kDa band observed is comparable to the molecular weight seen with other commercially available antibodies to ZO1 tight junction protein.
  • Overlay histogram showing MCF-7 cells stained with ab61357 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab61357, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References for Anti-ZO1 tight junction protein antibody [mAbcam 61357] (ab61357)

This product has been referenced in:

See 1 Publication for this product

Product Wall

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 (proximal colon)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: steamer also tried boiling, microwave
Permeabilization No
Specification proximal colon
Blocking step BSA/NDS as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Fixative Formaldehyde
Username

Miss. Anne Sailer

Verified customer

Submitted Aug 04 2015

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Colon)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: steamer also tried microwave and boiling
Permeabilization No
Specification Colon
Blocking step BSA as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Fixative Formaldehyde
Username

Miss. Anne Sailer

Verified customer

Submitted Aug 04 2015

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (16HBE human bronchiolar epithelium cell line)
Permeabilization Yes - triton X
Specification 16HBE human bronchiolar epithelium cell line
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
Fixative Paraformaldehyde
Username

Dr. David Johnston

Verified customer

Submitted Feb 17 2015

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Chicken Cell (neurulation stage chick embryos)
Permeabilization Yes - 0.3% TritonX-100 in PBS
Specification neurulation stage chick embryos
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative Paraformaldehyde
Username

Ms. Amanda Baumholtz

Verified customer

Submitted Apr 02 2014

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (brain tissue)
Permeabilization Yes - triton
Specification brain tissue
Blocking step Serum as blocking agent for 45 minute(s) · Concentration: 10µg/mL · Temperature: 25°C
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Apr 01 2014

The MCF7 FACS data we show on the datasheet came from cells that were harvested with trypsin, not from cells that were scraped from the culture dishes. If you are concerned that trypsin will remove ZO1 from your cells, I recommend comparing FACS data f...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"