ZO-1 Recombinant antibody

ZO-1 Uni-rAb^TM Recombinant Antibody for WB, IHC, IF/ICC, IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse

Applications

WB, IHC, IF/ICC, IP, ELISA

Conjugate

Unconjugated

CloneNo.

2P13

Cat no : 82870-1-RR

Synonyms

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

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Validation Data Gallery

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  WB analysis using 82870-1-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 82870-1-RR (ZO-1 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

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  IP experiment of HEK-293 using 82870-1-RR

  IP result of anti-ZO-1 (IP:82870-1-RR, 4ug; Detection:82870-1-RR 1:2000) with HEK-293 cells lysate 920 ug.

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  IHC staining of mouse kidney using 82870-1-RR

  Immunohistochemical analysis of paraffin-embedded mouse kidney tissue slide using 82870-1-RR (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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  IHC staining of mouse colon using 82870-1-RR

  Immunohistochemical analysis of paraffin-embedded mouse colon tissue slide using 82870-1-RR (ZO-1 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse colon using 82870-1-RR

  Immunohistochemical analysis of paraffin-embedded mouse colon tissue slide using 82870-1-RR (ZO-1 antibody) at dilution of 1:1000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of mouse colon using 82870-1-RR

  Immunofluorescent analysis of (4% PFA) fixed paraffin-embedded mouse colon tissue using ZO-1 antibody (82870-1-RR, Clone: 2P13 ) at dilution of 1:200 and Multi-rAb CoraLite ® Plus 488-Goat Anti-Rabbit Recombinant Secondary Antibody (H+L) (RGAR002). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of mouse heart using 82870-1-RR

  Immunofluorescent analysis of (4% PFA) fixed paraffin-embedded mouse heart tissue using ZO-1 antibody (82870-1-RR, Clone: 2P13 ) at dilution of 1:200 and Multi-rAb CoraLite ® Plus 488-Goat Anti-Rabbit Recombinant Secondary Antibody (H+L) (RGAR002). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of MCF-7 using 82870-1-RR

  Immunofluorescent analysis of (4% PFA) fixed MCF-7 cells using ZO-1 antibody (82870-1-RR, Clone: 2P13 ) at dilution of 1:1500 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 82870-1-RR

  Immunofluorescent analysis of (4% PFA) fixed MCF-7 cells using ZO1 antibody (82870-1-RR, Clone: 28567-1_2P13_230403 ) at dilution of 1:1000 and Multi-rAb CoraLite ® Plus 594-Goat Anti-Rabbit Recombinant Secondary Antibody (H+L) (RGAR004).

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  Affinity and Kinetic Characterization of 82870-1-RR

  Biolayer interferometry (BLl) kinetic assays of 82870-1-RR against Human ZO-1 were performed. The affinity constant is 0.723 nM.

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Tested Applications

• Positive WB detected in: Caco-2 cells, MCF-7 cells, HeLa cells, HepG2 cells
• Positive IP detected in: HEK-293 cells
• Positive IHC detected in: mouse kidney tissue, mouse colon tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: MCF-7 cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:10000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:1000-1:4000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:750-1:3000

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

82870-1-RR targets ZO-1 in WB, IHC, IF/ICC, IP, ELISA applications and shows reactivity with human, mouse samples.

• Tested Reactivity: human, mouse
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: ZO-1 fusion protein Ag33182
• Full Name: tight junction protein 1 (zona occludens 1)
• Calculated Molecular Weight: 1748 aa, 195 kDa
• Observed Molecular Weight: 230 kDa
• GenBank Accession Number: BC111712
• Gene Symbol: ZO-1
• Gene ID (NCBI): 7082
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Tight junction (or zonula occludens) forms the continuous intercellular barrier between epithelial and endothelial cells, which is required to separate tissue spaces and regulate the selective movement of solutes across the epithelium and endothelium (PMID: 20066090). ZO-1 (also known as TJP1) is a peripheral membrane phosphoprotein located on the cytoplasmic face and is expressed in tight junctions of both epithelial and endothelial cells (PMID: 3528172). It binds the transmembrane proteins occludin and the claudins linking them to cytoskeletal actin (PMID: 17418867). ZO-1 belongs to a family of multidomain proteins known as the membrane-associated guanylate kinase homologs (MAGUKs). It is a pivotal tight junction protein and may be involved in signaling mechanisms regulating cell proliferation and differentiation (PMID: 22782886).