Product nameAnti-ZO1 tight junction protein antibody [mAbcam 61357]
See all ZO1 tight junction protein primary antibodies
DescriptionMouse monoclonal [mAbcam 61357] to ZO1 tight junction protein
Tested applicationsSuitable for: Flow Cyt, WB, IPmore details
Species reactivityReacts with: Human
Predicted to work with: Mouse, Dog
Synthetic peptide corresponding to Human ZO1 tight junction protein aa 750-850 conjugated to keyhole limpet haemocyanin.
(Peptide available as
- This antibody gave a positive signal in the following whole cell lysates: HepG2; HEK293; A431; Caco2.
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 bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
Concentration information loading...
Clone numbermAbcam 61357
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.
|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.|
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.
modificationsPhosphorylated. 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
- Tight junction protein 1 antibody
- Tight junction protein ZO-1 antibody
- Tight junction protein ZO1 antibody
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.
All lanes : 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.
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
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.
This product has been referenced in:
- He L et al. Bradykinin potentially stimulates cell proliferation in rabbit corneal endothelial cells through the ZO-1/ZONAB pathway. Int J Mol Med 42:71-80 (2018). Read more (PubMed: 29568941) »
- Lv JW et al. Inhibition of microRNA-214 promotes epithelial-mesenchymal transition process and induces interstitial cystitis in postmenopausal women by upregulating Mfn2. Exp Mol Med 49:e357 (2017). WB . Read more (PubMed: 28729638) »