ZO-3, CT (Tight Junction-associated Polypeptide, Zonula Occludens 3)

ZO-3, a ~130kD protein, contains three PSD-95/SAP90, discs-large, ZO-1 (PDZ) domains, a SH3 domain, and a guanylate kinase-liked domain. Like ZO-1 and ZO-2, ZO-3 contains a C-terminal acidic domain and a basic region (between the first and second PDZ domains), making it belongs to the MAGUK protein family. By Western blot analysis, ZO-3 is identified as a ~130kD protein that directly interacts with tight junction proteins ZO-1 and occuldin, but not with ZO-2. However, in one study, ZO-2, ZO-3 and Occludin colocalized with actin aggregates at cell border in MDCK cells. ZO-3 is localized to sites of cell-cell interaction, identical to that of ZO-1 in tight junctions. By immunoprecipitation analysis, ZO-3 co-immunoprecipitates with ZO-1. Using MDCK cells, ZO-3 protein colocalized with ZO-1 by immunofluorescence. In vitro binding assays, the N-terminal of ZO-3 showed preferably binding of ZO-1 and actin filaments3. Moreover, the binding of ZO-1/ZO-2/ZO-3 was also found to interact with certain Claudin proteins.\n\nApplications: \nSuitable for use in Immunoflourescence, ELISA, Western Blot, Immunoprecipitation and Immunohistochemistry. Other applications not tested.\n\nRecommended Dilution:\nImmunofluorescence: 0.5-2ug/ml\nELISA: 0.1-1ug/ml \nWestern Blot: 1-3ug/ml \nImmunoprecipitation: 2-3ug\nImmunohistochemistry: 4-5ug/ml. Fixation of tissues with 2% paraformaldehyde/picric acid followed by NO post-fixation is required to produce staining in mouse liver tissues. Stronger fixation has been observed to abolish labeling.\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

Z0773

Size

100ug

Applications

ELISA, IF, IHC, IP, WB

Hosts

Rabbit

Reactivities

Hum, Mouse, Can

Form

Supplied as a liquid in PBS, pH 7.2, 0.1% sodium azide.

P Type

Pab

Purity

Purified by immunoaffinity chromatography.

Isotype

IgG

References

1. Haskins J, et al. J Cell Biol. 141(1):199-208, 1998. 2. Wittchen ES, et al. J Biol Chem. 274(49):35179-35185, 1999. 3. Wittchen ES, et al. J Cell Biol. 151(4):825-836, 2000. 4. Itoh M, et al. J Cell Biol. 147(6):1351-1363, 1999.\n

Additional Info

Recognizes the human ZO-3 protein. MW: ~130kD. Crossreactivity with other Zonula Occluden family members has not been observed. Reactivity is con?rmed with human T84, HeLa and canine MDCK cell lysates as well as mouse liver homogenates. Species Crossreactivity: canine and mouse.