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
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide. This information is considered to be commercially sensitive.

  • Positive control

    • WB: A549, HeLa, HeLa CCCP stimulated, PC-3, SK-OV-3, A431, HEK-293T and HEK-293T tunicamycin stimulated whole cell lysates. IHC-P: Mouse adipose tissue. ICC/IF: Caco-2, MCF7 cells.

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 long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab216880 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 1/1000. Detects a band of approximately 195, 187 kDa (predicted molecular weight: 195 kDa).
IHC-P 1/100 - 1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use a concentration of 1 - 5 µg/ml.

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

Images

  • All lanes : Anti-ZO1 tight junction protein antibody (ab216880) at 1/1000 dilution

    Lanes 1 & 15 : MW ladder (opal pre-stained)
    Lane 2 : WM-115 whole cell lysate
    Lane 3 : A549 whole cell lysate
    Lane 4 : HeLa whole cell lysate
    Lane 5 : HeLa CCCP stimulated whole cell lysate
    Lane 6 : PC-3 whole cell lysate
    Lane 7 : SK-OV-3 whole cell lysate
    Lane 8 : Mouse testis lysate
    Lane 9 : Rat testis lysate
    Lane 10 : A431 whole cell lysate
    Lane 11 : K562 whole cell lysate
    Lane 12 : HL-60 whole cell lysate
    Lane 13 : HEK-293T whole cell lysate
    Lane 14 : HEK-293T tunicamycin stimulated whole cell lysate

    Secondary
    All lanes : Rabbit secondary HRP antibody at 1/70000 dilution

    Developed using the ECL technique.

    Predicted band size: 195 kDa
    Observed band size: 187,195 kDa
    why is the actual band size different from the predicted?


    Exposure time: 11 seconds


    Blocking buffer: Blotto 5%.

    Dilution buffer: 1X PBS, 0.01% azide.

  • ab216880 stained in MCF7 (Human breast adenocarcinoma cell line) cells. Cells were fixed with 100% methanol (5 minutes) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 1 hour at room temperature to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab216880 at 1 µg/ml and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150120 (pseudo-colored red) and ab150081 (colored green) used at 1 µg/ml for 1 hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43 µM for 1 hour at room temperature

  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded mouse adipose tissue labeling ZO1 tight junction protein with ab216880 at 1/100 dilution (A) or 1/200 dilution (B). Visualized with WARP RED on MACH 4 universal AP polymer detection system.

  • Immunofluorescent analysis of 0.5% PFA-fixed (A and C) and 0.5% methanol-fixed (B and D) Caco-2 (human colorectal adenocarcinoma cell line) cells labeling ZO1 tight junction protein with ab216880 at 10 µg/ml (green).

    The nuclear counter stain is DAPI (blue).

    Secondary antibody only control: C and D.

References

This product has been referenced in:

  • Chen YF  et al. Inonotus obliquus polysaccharide ameliorates dextran sulphate sodium induced colitis involving modulation of Th1/Th2 and Th17/Treg balance. Artif Cells Nanomed Biotechnol 47:757-766 (2019). Read more (PubMed: 30856346) »
  • Luan L  et al. HOXD10 silencing suppresses human fibroblast-like synoviocyte migration in rheumatoid arthritis via downregulation of the p38/JNK pathway. Exp Ther Med 16:1621-1628 (2018). Read more (PubMed: 30186380) »
See all 4 Publications for this product

Customer reviews and Q&As

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cultured Cells (iPS cell derived intestinal cells)
Permeabilization
Yes - triton x-100
Specification
iPS cell derived intestinal cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

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Verified customer

Submitted Apr 26 2018

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