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.
|Tested species reactivity||Dog, Human|
|Published species reactivity||Dog, Human, Not Applicable|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Human recombinant ZO-1 fusion protein encompassing amino acids 334-634|
|Conjugate||Alexa Fluor® 488|
|Storage buffer||PBS, pH 7.4, with 1% BSA, 50% glycerol|
|Contains||0.1% sodium azide|
|Storage Conditions||4° C, store in dark|
|Tested Applications||Dilution *|
|ELISA (ELISA)||0.1-1.0 ug/ml|
|Immunocytochemistry (ICC)||5 µg/ml|
|Immunofluorescence (IF)||5 µg/ml|
|Western Blot (WB)||1-2 ug/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Tight junctions are complexes of proteins that create intercellular boundaries between the plasma membrane domains of epithelial and endothelial cells. Many of the tight junction-associated proteins are members of the membrane- associated guanylate kinase (MAGUK) family and include occludin, ZO-1, ZO-2 and ZO-3. These proteins are thought to have both structural and signaling roles, and are 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.
|Not Applicable||Not Cited||
A multi-stage process including transient polyploidization and EMT precedes the emergence of chemoresistent ovarian carcinoma cells with a dedifferentiated and pro-inflammatory secretory phenotype.
339188 was used in immunocytochemistry to study the multi-stage process ovarian carcinoma cells adopt upon becoming chemoresistant
|Rohnalter V,Roth K,Finkernagel F,Adhikary T,Obert J,Dorzweiler K,Bensberg M,Müller-Brüsselbach S,Müller R||Oncotarget (6:40005)||2015|
Cytoskeleton remodelling of confluent epithelial cells cultured on porous substrates.
339188 was used in immunocytochemistry to investigate the effect of substrate topography on cytoskeleton of epithelial cells
|Rother J,Büchsenschütz-Göbeler M,Nöding H,Steltenkamp S,Samwer K,Janshoff A||Journal of the Royal Society, Interface (12:null)||2015|
Thrombin induces epithelial-mesenchymal transition and collagen production by retinal pigment epithelial cells via autocrine PDGF-receptor signaling.
339188 was used in immunocytochemistry to investigate the effect of the coagulation proteins factor Xa and thrombin on the epithelial-mesenchymal transition and collagen production by RPE cells.
|Bastiaans J,van Meurs JC,van Holten-Neelen C,Nagtzaam NM,van Hagen PM,Chambers RC,Hooijkaas H,Dik WA||Investigative ophthalmology and visual science (54:8306)||2013|
Primordium of an artificial Bruch's membrane made of nanofibers for engineering of retinal pigment epithelium cell monolayers.
339188 was used in immunocytochemistry to test if nanofibrous membranes imitate the natural Bruch's membrane and allow for the engineering of an in vivo-like human retinal pigment epithelium.
|Warnke PH,Alamein M,Skabo S,Stephens S,Bourke R,Heiner P,Liu Q||Acta biomaterialia (9:9414)||2013|
rpS6 regulates blood-testis barrier dynamics through Akt-mediated effects on MMP-9.
339188 was used in immunohistochemistry - paraffin section to investigate the role of rpS6 in blood-testis barrier function
|Mok KW,Mruk DD,Cheng CY||Journal of cell science (127:4870)||2014|
Microtubule affinity-regulating kinase 4 (MARK4) is a component of the ectoplasmic specialization in the rat testis.
339188 was used in immunohistochemistry - frozen section to examine the role of actin and tubulin in ectoplasmic specialization
|Tang EI,Xiao X,Mruk DD,Qian XJ,Mok KW,Jenardhanan P,Lee WM,Mathur PP,Cheng CY||Spermatogenesis (2:117)||2012|