Anticorps Polyclonal de lapin anti-ZO-1

• Phare
• Validé par KD/KO

ZO-1 Polyclonal Antibody for WB, IHC, IF/ICC, IP, ELISA

Hôte / Isotype

Lapin / IgG

Réactivité testée

canin, Humain, rat, souris, Hamster et plus (5)

Applications

WB, IHC, IF/ICC, IP, CoIP, RIP, ELISA

Conjugaison

Non conjugué

Publications

1116

Avis (4)

N° de cat : 21773-1-AP

Synonymes

TJP1, ZO1, ZO 1, Tight junction protein ZO-1, Tight junction protein ZO 1

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Galerie de données de validation

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  WB analysis using 21773-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 21773-1-AP (ZO-1 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis using 21773-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 21773-1-AP (ZO-1 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

• 

  WB analysis using 21773-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 21773-1-AP (ZO-1 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  WB analysis of COLO 320 using 21773-1-AP

  COLO 320 cells were subjected to SDS PAGE followed by western blot with 21773-1-AP (ZO-1 antibody) at dilution of 1:800 incubated at room temperature for 1.5 hours.

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  IP experiment of HEK-293 using 21773-1-AP

  IP result of anti-ZO-1 (IP:21773-1-AP, 4ug; Detection:21773-1-AP 1:30000) with HEK-293 cells lysate 1800 ug.

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  IHC staining of mouse kidney using 21773-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse kidney tissue slide using 21773-1-AP (ZO-1 antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse kidney using 21773-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse kidney tissue slide using 21773-1-AP (ZO-1 antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of MCF-7 using 21773-1-AP

  Immunofluorescent analysis of (4% PFA) fixed MCF-7 cells using ZO-1 antibody (21773-1-AP) at dilution of 1:2000 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), Alpha Tubulin antibody (66031-1-Ig, Clone: 1E4C11, red).

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  IF Staining of MCF-7 using 21773-1-AP

  Immunofluorescent analysis of (-20°C Methanol) fixed MCF-7 cells using ZO-1 antibody (21773-1-AP) at dilution of 1:200 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L).

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  IF Staining of MDCK using 21773-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed MDCK cells using 21773-1-AP (ZO-1 antibody) at dilution of 1:600 and 66187-1-Ig (Cytokeratin 18 mouse mAb) at dilution of 1:400, further stained with CoraLite488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) for 21773-1-AP, and CoraLite594-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) for 66187-1-Ig.

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  IF Staining of MCF-7 using 21773-1-AP

  Immunofluorescent analysis of (4% PFA) fixed MCF-7 cells using ZO-1 antibody (21773-1-AP) at dilution of 1:2000 and CoraLite®594-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), Alpha Tubulin antibody (66031-1-Ig, Clone: 1E4C11, green).

• 

  IF Staining of HUVEC using 21773-1-AP

  Immunofluorescent analysis of (4% PFA) fixed HUVEC cells using ZO-1 antibody (21773-1-AP) at dilution of 1:2000 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L), Alpha Tubulin antibody (66031-1-Ig, Clone: 1E4C11, red).

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Applications testées

• Résultats positifs en WB: cellules A549, cellules Caco-2, cellules COLO 320, cellules HEK-293, cellules HeLa, cellules HepG2, cellules HT-29, cellules HUVEC, cellules K-562, cellules MCF-7, tissu cérébral de rat, tissu cérébral de souris
• Résultats positifs en IP: cellules HEK-293,
• Résultats positifs en IHC: tissu rénal de souris, ; il est suggéré de démasquer l'antigène avec un tampon de TE buffer pH 9.0; (*) À défaut, 'le démasquage de l'antigène peut être 'effectué avec un tampon citrate pH 6,0.
• Résultats positifs en IF/ICC: cellules MCF-7, cellules HUVEC, cellules MDCK

Dilution recommandée

• Western Blot (WB): WB : 1:5000-1:50000
• Immunoprécipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochimie (IHC): IHC : 1:1000-1:4000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:100-1:2000

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

Applications publiées

• KD/KO: See 3 publications below
• WB: See 796 publications below
• IHC: See 9 publications below
• IF: See 428 publications below
• ELISA: See 1 publications below
• FC: See 3 publications below
• CoIP: See 1 publications below
• RIP: See 1 publications below

Informations sur le produit

21773-1-AP cible ZO-1 dans les applications de WB, IHC, IF/ICC, IP, CoIP, RIP, ELISA et montre une réactivité avec des échantillons canin, Humain, rat, souris, Hamster

• Réactivité: canin, Humain, rat, souris, Hamster
• Réactivité citée: rat, bovin, canin, Chèvre, Humain, Lapin, porc, poulet, souris
• Hôte / Isotype: Lapin / IgG
• Clonalité: Polyclonal
• Type: Anticorps
• Immunogène: ZO-1 Protéine recombinante Ag16454
• Nom complet: tight junction protein 1 (zona occludens 1)
• Masse moléculaire calculée: 1748 aa, 195 kDa
• Poids moléculaire observé: 230 kDa
• Numéro d’acquisition GenBank: BC111712
• Symbole du gène: ZO-1
• Identification du gène (NCBI): 7082
• Conjugaison: Non conjugué
• Forme: Liquide
• Méthode de purification: Purification par affinité contre l'antigène
• Tampon de stockage: PBS avec azoture de sodium à 0,02 % et glycérol à 50 % pH 7,3
• Conditions de stockage: Stocker à -20°C. Stable pendant un an après l'expédition. L'aliquotage n'est pas nécessaire pour le stockage à -20^oC Les 20ul contiennent 0,1% de BSA.

Informations générales

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).  

Publications

Species	Application	Title
mouse		Am J Respir Crit Care Med; Ezrin, a Membrane Cytoskeleton Cross Linker Protein, as a Marker of Epithelial Damage in Asthma.; Authors - Man Jia
human,mouse	IF	Cell Metab; Gut microbiota modulate distal symmetric polyneuropathy in patients with diabetes; Authors - Junpeng Yang
human	IF	Cell Stem Cell; Human branching cholangiocyte organoids recapitulate functional bile duct formation.; Authors - Floris J M Roos
	IF	Microbiome; Arula-7 powder improves diarrhea and intestinal epithelial tight junction function associated with its regulation of intestinal flora in calves infected with pathogenic Escherichia coli O1; Authors - Hao Chen
mouse	IF	Adv Sci (Weinh); Plasma-Activated Solutions Mitigates DSS-Induced Colitis via Restoring Redox Homeostasis and Reversing Microbiota Dysbiosis; Authors - Tuanhe Sun
human	WB	J Extracell Vesicles; Extracellular vesicle-mediated delivery of circDYM alleviates CUS-induced depressive-like behaviours.; Authors - Xiaoyu Yu

Avis

The reviews below have been submitted by verified Proteintech customers who received an incentive forproviding their feedback.

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FH

Natalia (Verified Customer) (07-10-2024)

Used the dilution recommended (1:2000) and did not work. Haven't had the chance to re-try yet

• Applications: Immunofluorescence
• Primary Antibody Dilution: 1:2000
• Cell Tissue Type: Primary human endothelial cells

FH

Sarah (Verified Customer) (03-20-2024)

The antibody works perfect.

• Applications: Immunofluorescence
• Primary Antibody Dilution: 1:1000
• Cell Tissue Type: Cryosections intestine

ZO-1 Antibody Immunofluorescence validation (1:1000 dilution) in Cryosections intestine (Cat no:21773-1-AP)

FH

Xie (Verified Customer) (10-26-2021)

Efficient Labelling from proteintech ZO1 Antibody

• Applications: Immunofluorescence
• Primary Antibody Dilution: 1:150
• Cell Tissue Type: CaCo-2 cell line

ZO-1 Antibody Immunofluorescence validation (1:150 dilution) in CaCo-2 cell line (Cat no:21773-1-AP)

FH

Samuel (Verified Customer) (08-06-2021)

Clean and strong band at 220-230 kDa in WB (reducing and denaturing conditions). I used the antibody at a 1:2500 dilution in 5% milk, incubation overnight at 4 degrees Celsius. 30ug loaded per sample.

• Applications: Western Blot
• Primary Antibody Dilution: 1/2500
• Cell Tissue Type: SH-SY5Y, HEPG2, HEK293. 30ug each

ZO-1 Antibody Western Blot validation (1/2500 dilution) in SH-SY5Y, HEPG2, HEK293. 30ug each (Cat no:21773-1-AP)

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