Various lysates were subjected to SDS PAGE followed by western blot with 82870-7-RR (ZO1 antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.
Various lysates were subjected to SDS PAGE followed by western blot with 82870-7-RR (ZO1 antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.
WB analysis using 82870-7-RR
Various lysates were subjected to SDS PAGE followed by western blot with 82870-7-RR (ZO1 antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.
Various lysates were subjected to SDS PAGE followed by western blot with 82870-7-RR (ZO1 antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.
IP experiment of HEK-293 using 82870-7-RR
IP result of anti-ZO-1 (IP:82870-7-RR, 4ug; Detection:82870-7-RR 1:4000) with HEK-293 cells lysate 1610 ug.
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue slide using 82870-7-RR (ZO-1 antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IF Staining of MCF-7 using 82870-7-RR
Immunofluorescent analysis of (4% PFA) fixed MCF-7 cells using ZO-1 antibody (82870-7-RR, Clone: 240150D7 ) at dilution of 1:400 and CoraLite®594-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-4).
Immunofluorescent analysis of (4% PFA) fixed MCF-7 cells using ZO-1 antibody (82870-7-RR, Clone: 240150D7 ) at dilution of 1:400 and CoraLite®594-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-4).
IF Staining of U2OS using 82870-7-RR
Immunofluorescent analysis of (4% PFA) fixed U2OS cells using ZO-1 antibody (82870-7-RR, Clone: 240150D7 ) at dilution of 1:500 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2), Alpha Tubulin antibody (66031-1-Ig, Clone: 1E4C11, red).
Immunofluorescent analysis of (4% PFA) fixed U2OS cells using ZO-1 antibody (82870-7-RR, Clone: 240150D7 ) at dilution of 1:400 and Multi-rAb CoraLite ® Plus 488-Goat Anti-Rabbit Recombinant Secondary Antibody (H+L) (RGAR002).
FC experiment of MCF-7 using 82870-7-RR
1x10^6 MCF-7 cells were intracellularly stained with 0.25 ug ZO-1 Recombinant antibody (82870-7-RR, Clone:240150D7) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.25 ug Rabbit IgG Isotype Control Recombinant Antibody (98136-1-RR, Clone: 240953C9) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).
1x10^6 MCF-7 cells were intracellularly stained with 0.25 ug ZO-1 Recombinant antibody (82870-7-RR, Clone:240150D7) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.25 ug Rabbit IgG Isotype Control Recombinant Antibody (98136-1-RR, Clone: 240953C9) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).
Affinity and Kinetic Characterization of 82870-7-RR
Biolayer interferometry (BLl) kinetic assays of 82870-7-RR against Human ZO-1 were performed. The affinity constant is 16.2 nM.
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HeLa cells, mouse brain tissue, HepG2 cells, Caco-2 cells, HT-29 cells, MCF-7 cells, rat brain tissue
Positive IP detected in
HEK-293 cells
Positive IHC detected in
mouse kidney 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, U2OS cells, MDCK cells, Caco-2 cells
Positive FC (Intra) detected in
MCF-7 cells
Freshly prepared samples are recommended.
Recommended dilution
Application
Dilution
Western Blot (WB)
WB : 1:5000-1:50000
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 : 1:200-1:800
Flow Cytometry (FC) (INTRA)
FC (INTRA) : 0.25 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
82870-7-RR targets ZO-1 in WB, IHC, IF/ICC, FC (Intra), IP, ELISA applications and shows reactivity with human, mouse, rat, canine samples.
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 -20oC storage. 20ul sizes contain 0.1% BSA.
Background Information
Tight junction (or zonula occludens) form the continuous intercellular barrier between epithelial and endothelial cells, which is required to separate tissue spaces and regulate 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).
Various lysates were subjected to SDS PAGE followed by western blot with 82870-7-RR (ZO1 antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.
WB analysis using 82870-7-RR
Various lysates were subjected to SDS PAGE followed by western blot with 82870-7-RR (ZO1 antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours.
IHC Figures
IHC staining of mouse kidney using 82870-7-RR
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue slide using 82870-7-RR (ZO-1 antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).
IP Figures
IP experiment of HEK-293 using 82870-7-RR
IP result of anti-ZO-1 (IP:82870-7-RR, 4ug; Detection:82870-7-RR 1:4000) with HEK-293 cells lysate 1610 ug.
IF/ICC Figures
IF Staining of MCF-7 using 82870-7-RR
Immunofluorescent analysis of (4% PFA) fixed MCF-7 cells using ZO-1 antibody (82870-7-RR, Clone: 240150D7 ) at dilution of 1:400 and CoraLite®594-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-4).
IF Staining of U2OS using 82870-7-RR
Immunofluorescent analysis of (4% PFA) fixed U2OS cells using ZO-1 antibody (82870-7-RR, Clone: 240150D7 ) at dilution of 1:500 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2), Alpha Tubulin antibody (66031-1-Ig, Clone: 1E4C11, red).
IF Staining of Caco-2 using 82870-7-RR
Immunofluorescent analysis of (4% PFA) fixed Caco-2 cells using ZO-1 antibody (82870-7-RR, Clone: 240150D7 ) at dilution of 1:500 and Multi-rAb CoraLite ® Plus 488-Goat Anti-Rabbit Recombinant Secondary Antibody (H+L) (RGAR002), Alpha Tubulin antibody (66031-1-Ig, Clone: 1E4C11, red).
IF Staining of MDCK using 82870-7-RR
Immunofluorescent analysis of (4% PFA) fixed MDCK cells using ZO-1 antibody (82870-7-RR, Clone: 240150D7 ) at dilution of 1:500 and Multi-rAb CoraLite ® Plus 488-Goat Anti-Rabbit Recombinant Secondary Antibody (H+L) (RGAR002), CL594-Phalloidin (red).
IF Staining of U2OS using 82870-7-RR
Immunofluorescent analysis of (4% PFA) fixed U2OS cells using ZO-1 antibody (82870-7-RR, Clone: 240150D7 ) at dilution of 1:400 and Multi-rAb CoraLite ® Plus 488-Goat Anti-Rabbit Recombinant Secondary Antibody (H+L) (RGAR002).
FC (INTRA) Figures
FC experiment of MCF-7 using 82870-7-RR
1x10^6 MCF-7 cells were intracellularly stained with 0.25 ug ZO-1 Recombinant antibody (82870-7-RR, Clone:240150D7) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.25 ug Rabbit IgG Isotype Control Recombinant Antibody (98136-1-RR, Clone: 240953C9) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).
AFFINITY Figures
Affinity and Kinetic Characterization of 82870-7-RR
Biolayer interferometry (BLl) kinetic assays of 82870-7-RR against Human ZO-1 were performed. The affinity constant is 16.2 nM.
The species listed in Tested Reactivity are in-house verified and applicable species. For unlisted species, please refer to the homology analysis of the immunogen sequence and related species. For rabbit polyclonal antibodies, homology >70% is recommended. For mouse monoclonal antibodies and rabbit recombinant antibodies, homology >90% is recommended. Generally, the higher the homology, the greater the applicability. However, there will be certain differences in protein expression in different species, tissues or cells. Therefore, the homology analysis results are for reference only and do not serve as a guarantee.
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Proteintech
ZO-1 Recombinant antibody
Catalog Number
82870-7-RR
Citations
-
Dilutions
WB : 1:5000-1:50000 IP : 0.5-4.0 ug for IP and 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate for WB IHC : 1:1000-1:4000 IF/ICC : 1:200-1:800 FC (INTRA) : 0.25 ug per 10^6 cells in a 100 µl suspension
Applications
WB, IHC, IF/ICC, FC (Intra), IP, ELISA
Reactivity
human, mouse, rat, canine
Product Guarantee
Covers any species including not listed on datasheet
Covers any applications including not listed on datasheet
