Invitrogen

ZO-1 Monoclonal Antibody (ZO1-1A12), Alexa Fluor 488

Product Image Gallery

ZO-1 Antibody — Fig. 2 EPHB6 inhibits scattering of TNBC cells. a EPHB6-deficient TNBC cells, MDA-MB-231, were stably transfected with expression vectors encoding EPHB6 (MDA-B6), myc-tagged EPHB6 (MDA-B6-M), or mock-transfected with an empty vector (MDA-pc3). EPHB6 expression was analysed by Western blotting; tubulin and ERK2 represent loading controls. b MDA-pc3, MDA-B6 and MDA-B6-M cells were grown in individual colonies on glass coverslips in 24-well plates. Colonies were fixed and stained with phalloidin (red) and DAPI (blue). Representative colonies were imaged using an Olympus FV1000 confocal microscope with a 10x objective lens. Scale bar, 100 um, inserted using ImageJ. c Individual colonies of MDA-pc3, MDA-B6 and MDA-B6-M cells were fixed and stained with crystal violet in 6-well plates. Scattered and compact colonies were counted in six wells per cell line, using an inverted microscope. The graph represents the percentage of scattered colonies in each cell line. d TNBC cells, BT-20, were transduced with EPHB6-targeting shRNAs (shB6-1 or shB6-2, individually), as indicated. Transduction with non-silencing shRNA (BT20-NS) was used as a control. EPHB6 expression was analysed as in a , and quantitated by densitometry. EPHB6 quantifications were normalised on matching tubulin controls and presented in arbitrary units (AU). e Formation of scattered and compact colonies by BT20-NS, BT20-shB6-1 and BT20-shB6-2 cells was analysed as in c . f Representative confocal microscopy images of BT20

ZO-1 Antibody in Immunofluorescence (IF) — Immunofluorescence analysis of ZO-1 was performed using 90% confluent log phase Caco-2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with ZO-1 Monoclonal Antibody (ZO1-1A12), Alexa Fluor 488 at 5µg/mL in 0.1% BSA and incubated for 3 hours at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing junctional localization. Panel e shows the isotype control. The images were captured at 60X magnification.

Product Details

Tested Applications

Dilution

ELISA (ELISA)

0.1-1.0 µg/mL (unconjugated)

Immunocytochemistry (ICC)

5 µg/mL

Immunofluorescence (IF)

5 µg/mL

Western Blot (WB)

1-2 µg/mL

Published Applications

Immunofluorescence (IF)

See 4 publications below

Immunocytochemistry (ICC)

See 4 publications below

Immunohistochemistry (Paraffin) (IHC (P))

See 1 publication below

Western Blot (WB)

See 1 publication below

Immunohistochemistry (IHC)

See 1 publication below

Immunohistochemistry (Frozen) (IHC (F))

See 1 publication below

Product Specifications

Species Reactivity

Dog, Human

Published species

Dog, Human, Mouse

Host / Isotype

Mouse / IgG1

Class

Monoclonal

Type

Antibody

Clone

ZO1-1A12

Immunogen

Human recombinant ZO-1 fusion protein encompassing amino acids 334-634

Conjugate

Alexa Fluor® 488

Excitation/Emission Profile

View spectra

Form

Liquid

Concentration

0.5 mg/mL

Storage buffer

PBS, pH 7.4, with 4mg/mL BSA

Contains

0.1% sodium azide

Storage conditions

4° C, store in dark

RRID

AB_2532187

Product Specific Information

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. 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.

For Alexa Fluor 488 excitation 495 nm/emission 519 nm. Each lot of this antibody is tested for specificity by immunofluorescence. Expires one year from date of receipt when stored as instructed.

Target Information

ZO-1 forms complexes with either ZO-2 or ZO-3. In addition, these proteins can also associate with claudin, occludin and F-actin, at tight junction stands, where they provide a linkage between the actin cytoskeleton and the tight junction. ZO-1 expression is significantly reduced in many breast cancer lines. ZO-2 and ZO-3 are ubiquitously expressed within epithelial tight junctions, and unlike ZO-1, which is also expressed at cell junctions of cardiac myocytes, ZO-2 is not expressed in nonepithelial tissue. ZO-1 is a protein located on a cytoplasmic membrane surface of intercellular tight junctions. ZO-1 may be involved in signal transduction at cell-cell junctions. Two transcript variants encoding distinct isoforms have been identified for the ZO-1 gene. Diseases associated with ZO-1 dysfunction include Celiac Disease and Congenital Nephrotic Syndrome.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: Tight junction protein 1; tight junction protein 1 (zona occludens 1); Tight junction protein ZO-1; ZO-1; zona occludens 1; Zona occludens protein 1; zonula occludens 1 protein; Zonula occludens protein 1

Gene Aliases: TJP1; ZO-1; ZO1

UniProt ID: (Dog) O97758, (Human) Q07157

Entrez Gene ID: (Dog) 403752, (Human) 7082

Molecular Function: cell junction protein tight junction