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CoraLite®594-conjugated ZO-1 Polyclonal antibody

ZO-1 Polyclonal Antibody for FC (Intra), IF

Host / Isotype

Rabbit / IgG

Reactivity

human, Canine, mouse, rat, hamster

Applications

WB, IF, FC (Intra)

Conjugate

CoraLite®594 Fluorescent Dye

Cat no : CL594-21773

Synonyms

DKFZp686M05161, Tight junction protein 1, Tight junction protein ZO 1, TJP1, ZO 1, ZO1, ZO-1, Zona occludens protein 1, Zonula occludens protein 1



Tested Applications

Positive IF detected inMCF-7 cells, HUVEC cells
Positive FC detected inMCF-7 cells

Recommended dilution

ApplicationDilution
Immunofluorescence (IF)IF : 1:50-1:500
Flow Cytometry (FC)FC : 0.40 ug per 10^6 cells in a 100 µl suspension
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.
Sample-dependent, Check data in validation data gallery.

Product Information

CL594-21773 targets ZO-1 in WB, IF, FC (Intra) applications and shows reactivity with human, Canine, mouse, rat, hamster samples.

Tested Reactivity human, Canine, mouse, rat, hamster
Cited Reactivityhuman
Host / Isotype Rabbit / IgG
Class Polyclonal
Type Antibody
Immunogen ZO-1 fusion protein Ag16454
Full Name tight junction protein 1 (zona occludens 1)
Calculated Molecular Weight 1748 aa, 195 kDa
Observed Molecular Weight 230 kDa
GenBank Accession NumberBC111712
Gene Symbol TJP1
Gene ID (NCBI) 7082
RRIDAB_2919863
Conjugate CoraLite®594 Fluorescent Dye
Excitation/Emission Maxima Wavelengths588 nm / 604 nm
Form Liquid
Purification MethodAntigen affinity purification
Storage Buffer PBS with 50% Glycerol, 0.05% Proclin300, 0.5% BSA, pH 7.3.
Storage ConditionsStore at -20°C. Avoid exposure to light. Stable for one year after shipment. Aliquoting is unnecessary for -20oC storage. 20ul sizes contain 0.1% BSA.

Background Information

What is the function of ZO-1?

Zona Occludens 1 (ZO-1) is a tight junction (TJ) protein found in complexes at cell-cell contacts. The role of ZO-1 is to recruit other TJ proteins.1,2 The resulting TJ complexes regulate paracellular flow, contribute to apical-basal polarity, and are part of signaling pathways for proliferation and differentiation.3 This protein is useful for highlighting cell-cell contacts both in cultured cells and in tissue samples, outlining cell membranes and specific structures.

 

Where is ZO-1 expressed?

The protein is localized to the cytoplasmic membrane in most types of cells, particularly where a barrier function is essential. It is common in endothelial cells,4 which line the internal surface of blood and lymph vessels, and in epithelial cells,5 which form the outer barrier of organs.

ZO-1 is expressed in many tissues including the intestine, kidney, liver, and skeletal muscle cells.

 

What proteins does ZO-1 interact with?

The molecular weight of ZO-1 is 220kDa6 and it contains multiple distinct protein domains that allow bind to other junctional proteins at the cytoplasmic membrane. The N-terminal of ZO-1 can dimerize with other ZO proteins and bind directly to other TJ proteins including claudins, connexions, and JAMs,7-10 which allows it to assemble TJ complexes. The C-terminal can interact with actin and cortactin, anchoring the TJ complexes to the cytoskeleton.8,11 This suggests that ZO-1 forms a link between the outer cell-cell contacts and the inner actin.

 

1.     McNeil, E., Capaldo, C. T. & Macara, I. G. Zonula occludens-1 function in the assembly of tight junctions in Madin-Darby canine kidney epithelial cells. Mol. Biol. Cell 17, 1922-32 (2006).

2.     Kratzer, I. et al. Complexity and developmental changes in the expression pattern of claudins at the blood-CSF barrier. Histochem. Cell Biol. (2012). doi:10.1007/s00418-012-1001-9

3.     Guillemot, L., Paschoud, S., Pulimeno, P., Foglia, A. & Citi, S. The cytoplasmic plaque of tight junctions: A scaffolding and signalling center. Biochim. Biophys. Acta - Biomembr. 1778, 601-613 (2008).

4.     Tornavaca, O. et al. ZO-1 controls endothelial adherens junctions, cell-cell tension, angiogenesis, and barrier formation. J. Cell Biol. 208, 821-38 (2015).

5.     Umeda, K. et al. Establishment and characterization of cultured epithelial cells lacking expression of ZO-1. J. Biol. Chem. 279, 44785-94 (2004).

6.     Stevenson, B. R. Identification of ZO-1: a high molecular weight polypeptide associated with the tight junction (zonula occludens) in a variety of epithelia. J. Cell Biol. 103, 755-766 (1986).

7.     Itoh, M. et al. Direct Binding of Three Tight Junction-Associated Maguks, Zo-1, Zo-2, and Zo-3, with the Cooh Termini of Claudins. J. Cell Biol. 147, 1351-1363 (1999).

8.     Fanning, A. S., Jameson, B. J., Jesaitis, L. A. & Anderson, J. M. The Tight Junction Protein ZO-1 Establishes a Link between the Transmembrane Protein Occludin and the Actin Cytoskeleton. J. Biol. Chem. 273, 29745-29753 (1998).

9.     Kausalya, P. J., Reichert, M. & Hunziker, W. Connexin45 directly binds to ZO-1 and localizes to the tight junction region in epithelial MDCK cells. FEBS Lett. 505, 92-96 (2001).

10. Itoh, M. et al. Junctional adhesion molecule (JAM) binds to PAR-3. J. Cell Biol. 154, 491-498 (2001).

11. Itoh, M., Nagafuchi, A., Moroi, S. & Tsukita, S. Involvement of ZO-1 in Cadherin-based Cell Adhesion through Its Direct Binding to α Catenin and Actin Filaments. J. Cell Biol. 138, 181-192 (1997).  

Protocols

Product Specific Protocols
IF protocol for CL594 ZO-1 antibody CL594-21773Download protocol
Standard Protocols
Click here to view our Standard Protocols

Publications

SpeciesApplicationTitle
humanWB,IF

Chin J Nat Med

Stigmasterol protects human brain microvessel endothelial cells against ischemia-reperfusion injury through suppressing EPHA2 phosphorylation

Authors - Suping Li
humanIF

J Ginseng Res

Ginsenoside Rd protects cerebral endothelial cells from oxygen-glucose deprivation/reoxygenation induced pyroptosis via inhibiting SLC5A1 mediated sodium influx

Authors - Suping Li