- 36-4000 - Invitrogen Antibodies TJP3 antibody

Submitted references E-cadherin is downregulated in benign prostatic hyperplasia and required for tight junction formation and permeability barrier in the prostatic epithelial cell monolayer.

Li F, Pascal LE, Stolz DB, Wang K, Zhou Y, Chen W, Xu Y, Chen Y, Dhir R, Parwani AV, Nelson JB, DeFranco DB, Yoshimura N, Balasubramani GK, Gingrich JR, Maranchie JK, Jacobs BL, Davies BJ, Hrebinko RL, Bigley JD, McBride D, Guo P, He D, Wang Z
The Prostate 2019 Aug;79(11):1226-1237

Family-wide investigation of PDZ domain-mediated protein-protein interactions implicates β-catenin in maintaining the integrity of tight junctions.

Gujral TS, Karp ES, Chan M, Chang BH, MacBeath G
Chemistry & biology 2013 Jun 20;20(6):816-27

Delineation of the key aspects in the regulation of epithelial monolayer formation.

Aschauer L, Gruber LN, Pfaller W, Limonciel A, Athersuch TJ, Cavill R, Khan A, Gstraunthaler G, Grillari J, Grillari R, Hewitt P, Leonard MO, Wilmes A, Jennings P
Molecular and cellular biology 2013 Jul;33(13):2535-50

Macrophages and dendritic cells express tight junction proteins and exchange particles in an in vitro model of the human airway wall.

Blank F, Wehrli M, Lehmann A, Baum O, Gehr P, von Garnier C, Rothen-Rutishauser BM
Immunobiology 2011 Jan-Feb;216(1-2):86-95

Connexin47, connexin29 and connexin32 co-expression in oligodendrocytes and Cx47 association with zonula occludens-1 (ZO-1) in mouse brain.

Li X, Ionescu AV, Lynn BD, Lu S, Kamasawa N, Morita M, Davidson KG, Yasumura T, Rash JE, Nagy JI
Neuroscience 2004;126(3):611-30

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Lane 1 was probed with Rb anti-ZO-3 (Mid) (Product # 36-4000) and Lane 2 was probed with Ms anti-ZO-1 (Product # 33-9100).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Zona Occludens protein 3 (ZO-3) was performed by loading 50 µg of MDCK (lane 2) cell lysates and 2 µL SeeBlue® Plus2 Prestained Protein Ladder (Product # LC5925) (lane 1) per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane and blocked with 1% BSA/TBST for at least 1 hour at room temperature. ZO-3 was detected using a polyclonal antibody (Product # 36-4000) at a concentration of 1 µg/mL in blocking buffer overnight at 4°C on a rocking platform, followed by a goat anti-rabbit IgG Alexa Fluor 680 conjugated secondary antibody (Product # A-21109) at a dilution of 1:10,000 for at least 1 hour. Fluorescent detection was performed using the Odyssey® CLx imaging system (Li-cor Biosciences). Images generated by Joell Solan in Paul Lampe Lab at Fred Hutchinson Cancer Research Center.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of ZO-3 was performed by loading 30 µg of Caco-2 (lane1) and MCF7 (lane2) cell lysate using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800), and Pierce™ Power Blotter System (22834). Proteins were transferred to a nitrocellulose membrane and blocked with 5 % skim milk for 1 hour at room temperature. ZO-3 was detected at ~ 130 kDa using ZO-3 Rabbit Polyclonal Antibody (Product # 36-4000) at 1-2 µg/mL in 5 % skim milk at 4°C overnight on a rocking platform on a rocking platform. Goat Anti-Rabbit IgG - HRP Secondary Antibody (G21234) at 1:5000 dilution was used and chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of ZO-3/TJP3 Antibody was done on 90% confluent log phase CACO2 cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with ZO-3/TJP3 Antibody (Product # 36-4000) at 1µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor 488 Goat Anti-Rabbit IgG Secondary Antibody (Product # A-11008) at a dilution of 1:400 for 45 minutes 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 Alexa Fluor 594 Phalloidin (Product # A12381). Panel d is a merged image showing cell junctional localization. Panel e is a no primary antibody control. The images were captured at 40X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of ZO-3/TJP3 showing staining in the membrane of paraffin-embedded human colon tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a ZO-3/TJP3 polyclonal antibody (Product # 36-4000) diluted in 3% BSA-PBS at a dilution of 1:100 overnight at 4ºC in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of ZO-3/TJP3 showing staining in the membrane of paraffin-embedded human kidney tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a ZO-3/TJP3 polyclonal antibody (Product # 36-4000) diluted in 3% BSA-PBS at a dilution of 1:100 overnight at 4ºC in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

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

Supportive validation

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

36-4000

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