Invitrogen

ZO-3 Recombinant Polyclonal Antibody (24HCLC)

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ZO-3 Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence was performed on fixed and permeabilized Caco-2 cells for detection of ZO-3 using Anti-ZO-3 Recombinant Rabbit Polyclonal Antibody (Product # 710857, 1 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of ZO-3 protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating junctional localization of ZO-3. Panel e) represents control cells with no primary Antibody to assess background.

ZO-3 Antibody in Western Blot (WB) — Western blot analysis was performed on whole cell extracts (30 µg lysate) of Caco-2 (Lane1), MCF-7 (Lane 2) and HT-29 (Lane 3). The blots were probed with ZO-3 Recombinant Rabbit Polyclonal Antibody (Product # 710857, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 140 kDa band corresponding to ZO-3 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002), and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Product Details

710857

Applications	Tested Dilution	Publications
Western Blot (WB)	1-2 µg/mL	View 1 publication 1 publication
Immunocytochemistry (ICC/IF)	1 µg/mL	-

Product Specifications
Species Reactivity	Human
Published species	Human
Host/Isotype	Rabbit / IgG
Class	Recombinant Polyclonal
Type	Antibody
Clone	24HCLC
Immunogen	Protein corresponding to Human ZO-3 (aa 700-909) View immunogen
Conjugate	Unconjugated
Form	Liquid
Concentration	0.5 mg/mL
Purification	Protein A
Storage buffer	PBS, pH 7.2
Contains	0.09% sodium azide
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_2609947

Product Specific Information

This antibody is predicted to react with Monkey, Pig and Bovine.

Recombinant rabbit polyclonal antibodies are unique offerings from Thermo Fisher Scientific. They are comprised of a selection of multiple different recombinant monoclonal antibodies, providing the best of both worlds - the sensitivity of polyclonal antibodies with the specificity of monoclonal antibodies - all delivered with the consistency only found in a recombinant antibody. While functionally the same as a polyclonal antibody - recognizing multiple epitope sites on the target and producing higher detection sensitivity for low abundance targets - a recombinant rabbit polyclonal antibody has a known mixture of light and heavy chains. The exact population can be produced in every lot, circumventing the biological variability typically associated with polyclonal antibody production.

Target Information

ZO-3 is a member of the membrane associated guanylate kinse (MAGUK) family and are considered tight junction-associated proteins. Tight junctions are complexes of proteins that create intercellular boundaries between the plasma membrane domains of epithelial and endothelial cells. ZO-3 is believed to have both structural and signaling roles, and is characteristically defined by three protein-protein interaction modules: the PDZ domain, the SH3 domain and the guanylate kinase (GuK) domain. 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.

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

Product References

Long non‑coding RNA NEAT1 promotes ovarian cancer cell invasion and migration by interacting with miR‑1321 and regulating tight junction protein 3 expression.

Molecular medicine reports

Luo M,Zhang L,Yang H,Luo K,Qing C

710857 was used in Western Blotting to demonstrate the function and working mechanism of lncRNA nuclear enriched abundant transcript 1 (NEAT1) in ovarian cancer (OC).

Thu Oct 01 00:00:00 EDT 2020

Long non‑coding RNA NEAT1 promotes ovarian cancer cell invasion and migration by interacting with miR‑1321 and regulating tight junction protein 3 expression.

Molecular medicine reports

Luo M,Zhang L,Yang H,Luo K,Qing C

710857 was used in Western Blotting to demonstrate the function and working mechanism of lncRNA nuclear enriched abundant transcript 1 (NEAT1) in ovarian cancer (OC).

Thu Oct 01 00:00:00 EDT 2020

Bioinformatics

Protein Aliases: Tight junction protein 3; tight junction protein 3 (zona occludens 3); Tight junction protein ZO-3; TJP-3; Zona occludens protein 3; Zonula occludens protein 3

Gene Aliases: TJP3; ZO-3; ZO3

UniProt ID: (Human) O95049

Entrez Gene ID: (Human) 27134

Molecular Function: tight junction

Process(es)

protein binding

ZO-3 Recombinant Polyclonal Antibody (24HCLC)

Product Details

710857

Western Blot (WB)

Immunocytochemistry (ICC/IF)

Species Reactivity

Published species

Host/Isotype

Class

Type

Clone

Immunogen

Conjugate

Form

Concentration

Purification

Storage buffer

Contains

Storage conditions

RRID

Product Specific Information

Target Information

Bioinformatics

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