33-9111

antibody from Invitrogen Antibodies

Targeting: TJP1 DKFZp686M05161, MGC133289, ZO-1

Provider product page for 33-9111

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Antibody data

Antibody Data
Antigen structure
References [35]
Comments [0]
Validations
Immunocytochemistry [1]
Other assay [31]

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Product number: 33-9111 - Provider product page
Provider: Invitrogen Antibodies
Product name: ZO-1 Monoclonal Antibody (ZO1-1A12), FITC
Antibody type: Monoclonal
Antigen: Recombinant full-length protein
Description: Species reactivity includes human (Caco-2 Cell line) and dog (MDCK cell line). Based on sequence homology, reactivity with other species is likely but has not been confirmed. This monoclonal antibody detects ZO-1alpha+ and ZO-1alpha- isoforms. The antibody is specific for ZO-1 proteins; cross-reactivity with the related ZO-2 protein has not been observed. For this FITC conjugate the excitation (EX)/emission (EM) are 494/520 (nm).
Reactivity: Human, Canine
Host: Mouse
Conjugate: Green dye
Isotype: IgG
Antibody clone number: ZO1-1A12
Vial size: 100 µg
Concentration: 0.5 mg/mL
Storage: 4° C, store in dark

TJP1 protein structure - 33-9111 shown in red.

Supportive validation

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Experimental details: NULL
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 4 Inhibition of zonula occludens type 1 (ZO-1) expression in CBA/J/Hsd mouse bladders following 14 days of treatment with as -APF. Bladder sections from mice treated with as -APF following bladder epithelial injury show decreased ZO-1 immunofluorescence staining (shown by small junctions between cells indicated by arrows) as compared to mice treated with PBS or inactive control nonglycosylated peptide. Representative data shown for the 3 mice in each treatment group from one experiment; experiment performed three times. (500X final magnification).
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5. Effects of TNFalpha on protein distribution within Sertoli cells. Sertoli cells (0.04 x 10 6 cells/cm 2 ) were cultured on Matrigel(tm)-coated micro cover glasses for 4 d and treated with TNFalpha (25 ng/ml) for 24 h as described in Materials and Methods. Sertoli cells were then dual-labeled for occludin (red)/ZO-1 (green), or labeled for F-actin or cortactin (both green). Corresponding images were merged to show areas of co-localization (orange, A ). Nuclei were visualized with DAPI (blue, A and B ). Scale bars, 20 mum.
Conjugate: Green dye

Supportive validation

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Experimental details: Figure 2 Fb1 knock-down increases cell fusion in syn1 expressing cells. Fb1 -specific mRNA knock-down in BeWo choriocarcinoma cells using distinct siRNA (siFb1a and siFb1b) or their respective scrambled siRNA controls. Fb1 knock down induced a significant increase in cell fusion (a, d). (a) ZO-1-FITC and Hoechst 33342 immunocytochemistry was performed 72 hours after siRNA exposure and images are depicted in the lower row. The upper row shows matched, phase-contrast images. Scale bar represents 100 mum. Quantitative RT-PCR, immunoprecipitation and immunoblotting confirm decreases in (b) Fb1 mRNA (50-60% knock-down) and (c) protein (cell-associated and secreted) in siRNA treated Bewo cells. Expression (b) is normalized to unexposed samples cultured for similar time periods (48 or 72H). (d) Mean fusion indices for siRNA-exposed and control cells (error bars represent standard deviations; n = 4 x 5 fields). Cell fusion was assessed using phase contrast microscopy and quantitated using cell fusion indices. All observations were performed at a final magnification of 200x and total nuclei were counted per field using Leica MetaMorph image analyzing software. The number of fused syncytial aggregates and the number of nuclei in each aggregate was counted manually and fusion indices were defined as [(N-S)/T] x 100. N is the number of nuclei in syncytia, S is the number of syncytia, and T is the total number of nuclei counted. The fusion index quantitates the percentage of fusion events
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 MZ oep mutants have aberrant neural tube organization. (A) Projection of confocal z-series showing dorsal view of brain and anterior spinal cord. The midline ventricle is lined by ZO-1 expression (green) and the basal regions of the neuroepithelium are lined by the GFAP expression (red). (B) Projection of confocal z-series showing dorsal view of brain and anterior spinal cord from MZ oep mutant. Both apical ZO-1 and basal GFAP expression show extensive disruption to neural tube morphology in brain regions (arrowheads) but appear relatively normal in anterior spinal cord (arrow). (C) Projection of confocal z-series showing dorsal view of brain and anterior spinal cord from embryo treated with the Nodal inhibitor SB-431542. (D,E) Transverse sections show the normal single midline domain of ZO-1 appears discontinuous and more randomly oriented in MZ oep embryo. (F,G) The basal marker GFAP is expressed in ectopic foci deep from the surface of the MZ oep neural primordium. (H,I,J,K) Neurons labeled with tg(HUC-GFP) (green) and their axons labeled with Ac-tub (red) are present but disorganized in the Nodal-defective embryo brains. (L) By 28 hpf ectopic ventricles (arrowed) have opened up in the MZ oep brains. Ac-tub, anti-acetylated tubulin antibody; GFAP, glial fibrillary acidic protein; hpf, hours post fertilization; MZ oep , maternal-zygotic one-eyed pinhead ; ZO-1, zonula occludens 1.
Conjugate: Green dye

Supportive validation

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Experimental details: Figure 2 Ectopic divisions do not generate the abnormal neural tube in MZ oep embryos. (A) Location and orientation of divisions monitored over a 2-hour time-lapse period of neural rod development in wild-type embryo by analyzing the expression of apical marker Pard3-GFP (green) and H2B-RFP mRNA to label nuclei (not included in image for clarity, red dumbbells indicate location and orientation of dividing cells). Wild-type neural progenitor divisions are strongly orientated along the mediolateral axis of the developing neural tube. Yellow dots outline the rod. (B) Location and orientation of divisions monitored over a 2-hour time-lapse period of neural rod development in MZ oep embryo. (C,D) Orientation plots of divisions in wild-type and MZ oep embryos. (E,F,G) Dorsal projections of ventricle morphology (ZO-1, green) at 24 hpf in wild-type, MZ oep embryos and MZ oep embryos treated with CDIs (hydroxyurea and aphidicolin). Blocking division does not rescue ventricle morphology. PH3 staining of mitotic figures (purple) used to calculate efficiency of division block. (H) Transverse section of brain from MZ oep embryo treated with division blockers. (I) Quantification of divisions in wild-type, MZ oep and MZ oep division blocked embryos. Number of cell divisions between wild-type 194 and MZ oep 206, P = 0.5129 Student's t -test and between MZ oep 206 and MZ oep + CDI 39, *** P
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 4 Loss of mesoderm, but not Nodal signaling, leads to neural tube defects. (A,B,C,D,E,F,G) Projection of confocal z-series showing dorsal view of 24 hpf zebrafish embryos staining for the apical marker ZO-1 (green). (A) Disrupted ventricular organization of embryo treated with the Nodal inhibitor SB-431542 from the one-cell stage. (B) Treatment with SB-431542 from 70% epiboly does not cause neural tube defects. (C,D) Ventricle organization in Nodal mutants cyc and oep is largely normal. (E,F) Ventricle organization in ntl and syu mutants is normal. (G) Ventricle organization in the endoderm mutant cas mutants is normal. (H) Expression of the Nodal-dependent markers lefty and pitx2 show the SB-431542 drug is an efficient blocker of Nodal signaling. All embryos between 21 and 23 hpf, dorsal view. Arrowhead in (H) indicates asymmetric lefty expression at 21 hpf. (I,I') Two frames from time-lapse sequence show transplanted MZ oep cells (green) integrate and divide to make mirror-image daughters just like host cells in a wild-type embryo at 15 hpf. (J,J') Two frames at 15 hpf from time-lapse sequence show wild-type cells (green) divide to make mirror-image daughters after transplantation to a MZ oep embryo. Unlike divisions in wild-type embryos, however, these divisions can be far from their normal location at the midline. hpf, hours post fertilization; MZ oep , maternal-zygotic one-eyed pinhead ; wt, wild-type; ZO-1, zonula occludens 1.
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 Mesoderm rescues neural tube morphogenesis in MZ oep embryos. (A) Lateral view of an MZ oep embryo previously injected with the activated form of the TGF-beta receptor Taram-A* (Tar*) into a single blastomere. Rescued head mesoderm expresses GFP (green). (B) Transverse section of Taram-A*-injected embryos show rescued mesoderm (green and arrowed) underlying neural plate at 13 hpf. (C) By 24 hpf rescued mesoderm almost surrounds the neural tube (nt) in MZ oep embryos. The morphology of the neural tube is rescued and contains a single well-defined midline lumen (black arrow). (D) Rescued neural lumen morphology revealed by ZO-1 expression. (E,F) Identity, distribution and presence of rescued mesoderm, confirmed by the mesendoderm marker foxc1a , in wild-type and Taram-A*-injected MZ oep embryos. GFP, green fluorescent protein; hpf, hours post fertilization; MZ oep , maternal-zygotic one-eyed pinhead ; TGF-beta, transforming growth factor beta; ZO-1, zonula occludens 1.
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 9 Apical polarity development in MZ oep . ZO-1 staining in (A,B,C,D) wild-type and (E,F,G,H) MZ oep embryos at different stages of neurulation. All images are maximum confocal projections of six consecutive z-levels taken from a dorsal view at posterior hindbrain and anterior spinal cord regions. Anterior is up. White arrows indicate the dorsal midline of the embryo. The ZO-1 staining surrounding the edge of the neural tissue is from the polarized enveloping layer overlying the neural tissue. Scale bar is 50 mum. (I) The average number of pixels of ZO-1 staining over time for MZ oep and wild-type embryos. At 13.5 hpf the number of ZO-1 puncta in MZ oep embryos is significantly different to wild-type (* P
Conjugate: Green dye

Supportive validation

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Experimental details: Fig. 1 Defects in junctional remodelling upon inactivation of PI3Kalpha in endothelial cells. a Lateral views of intersomitic vessels (ISV) in vehicle and GDC-0326 (50 muM)-treated transgenic Tg( kdrl:EGFP ) s843 (shown in red) embryos stained for ZO-1 (green) at 33 h post fertilization (hpf). Single ZO-1 staining is shown in upper row. DA refers to dorsal aorta and DLAV refers to dorsal longitudinal anastomotic vessels. White lines indicate ZO-1 negative staining; punctuate white lines indicate elongation of junction; yellow arrowheads show ring-shape junctions. b Quantification of the length of the dorsal part of the ISVs without ZO-1 (left graph) and length of the ISVs with continuous ZO-1 staining (right graph) in vehicle and GDC-0326 treated embryos ( n >= 54 ISVs per treatment). c Representative maximum intensity projections of anti-VE-cadherin (green) and isolectin B4 (IB4, red) immunostained control and Pik3ca KD/iDeltaEC mouse retinas at P7. Single channel is shown in upper row. Yellow islets show higher magnification of selected regions shown to the right. Yellow arrowheads indicate vascular segments without VE-cadherin staining; red asterisks indicate VE-cadherin-positive isolated rings or single-dots within the vascular tubes, indicating cell-cell junctional contacts that have not elongated. d , e Quantitative number of VE-cadherin-negative vessels (junctional gaps) per unit area ( n >= 5 retinas per genotype) ( d ) and length of vessel structures wi
Conjugate: Green dye

Supportive validation

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Experimental details: Figure 3 Rai14 is an apical and basal ES protein in the rat testis. Dual-labeled immunofluorescence analysis was used to assess co-localization of Rai14 (red) with constituent protein laminin-gamma3 chain (green), actin regulatory protein Arp3 (green), actin binding protein drebrin E (green) and actin filament cross-linking protein palladin (green) at the apical ES in stage VII tubules when these proteins were all highly expressed. It was found that Rai14 indeed partially co-localized with each of these apical ES proteins (see ""orange yellow"" in merged images) as shown in ( A ). Rai14 (red) also partially co-localized with basal ES/TJ protein ZO-1 (see ""yellow"" arrowheads), and to a lesser extent with N-cadherin (see ""white"" arrowheads) in stage IV-V tubules when these proteins were highly expressed as shown in ( B ). Bar = 10 um in ( A ) or ( B ), which applies all other images in ( A ) and ( B ).
Conjugate: Green dye

Supportive validation

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Experimental details: Immunostaining of Cells in the Rocked Device, Multi-cell line Static Function Control, and Single-cell line Static Function Controls. Fluorescent confocal images of the nonbarrier cell lines (A549 and Caco2) were observed after immunostaining for surfactant (red) and tight junctions (protein ZO-1, green). The nuclei were counter-stained (DAPI, blue). The cultures were observed under a water immersion 25x collared objective and further magnified with the Zen program. Scale bars are present on the micrographs.
Conjugate: Green dye

Supportive validation

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Experimental details: 1 FIGURE Depletion of MDSC accelerates corneal allograft rejection. (A) Graphic illustration of experimental design. Isogeneic (BALB/c to BALB/c) and allogeneic (C57BL/6 to BALB/c) corneal transplantation was performed on day 0. Anti-Gr-1 antibody or control IgG was interperitoneally administered twice weekly for 6 weeks after transplantation. (B) Kaplan-Meier survival curves of isografts ( n = 5) and allografts ( n = 10 each group). **, P < .01; ns, not significant; log-rank tests. (C) Representative slim-lamp photographs (front view) of graft-bearing eyeballs on day 7 and day 15 posttransplant. Dashed circles outline corneal grafts. White arrows and arrowheads denote representative neovessels in the corneal graft and the graft bed, respectively. Open arrowheads in photographs of the isograft denote representative blood vessels in the iris. (D) H&E stain of corneal grafts on day 15 posttransplant from mice treated as in (A-C). Scale bars, 200 mum. (E) Representative slim-lamp photographs (front view and profile view) of normal (graft-free) eyeballs from mice treated with anti-Gr-1 antibody or PBS control on day 15 posttransplant as in (C). (F) Representative immunohistochemistry staining of the tight junction protein zonula occludens-1 (ZO-1) and TUNEL staining of corneal endothelium of normal eyeballs from untreated mice or mice treated with anti-Gr-1 antibody on day 7 and day 15 posttransplant as in (A-C). Nuclei of apoptotic cells were stained red in TUNEL assay and DNase
Conjugate: Green dye

Supportive validation

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Experimental details: 10.1371/journal.pone.0246115.g006 Fig 6 Alterations of the retinal pigment epithelium after suturing of the vortex veins. (A) Immunohistochemistry using anti-ZO-1 antibody for RPE flatmount in the control C57BL/6 mouse eye shows well organized and packed RPE cells. (B) 7 days after the vortex vein suturing, focal RPE cell degeneration is observed. Scale bar: 50mum.
Conjugate: Green dye

Supportive validation

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Main image
Experimental details: Figure 6 Immunohistochemistry analysis of retinal ZO-1. Primary antibody to ZO-1 (1:1,000) followed by secondary antibody Alexa Fluor donkey anti-rabbit 488 (1:500) identifies immunohistochemical staining in mouse retina sections at 1 month ( A - D ) and 3 months ( E - H ) localized to the OLM. A, C, E, G : The OLM in the B6J background retinas is unbroken. B, D, F, H : The OLM in the B6N background retinas is discontinuous. The top-right corners show higher magnification images. Lower magnification images were taken with a 20X objective lens (scale bar, 50 um); higher magnification images were taken at 100X objective (scale bar, 20 um). OLM=outer limiting membrane; ONL=outer nuclear layer.
Conjugate: Green dye