Technical Data

Z0774
Clone Type
Polyclonal
Host
Rabbit
Source
Human
Isotype
IgG
Grade
Affinity Purified
Applications
E IF IP WB
Crossreactivity
Ca Hu
Shipping Temp
Blue Ice
Storage Temp
-20°C
Rabbit Anti-ZO-3, Mid (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 filaments. Moreover, the binding of ZO-1/ZO-2/ZO-3 was also found to interact with certain Claudin proteins.

Applications
Suitable for use in Immunofluorescence, ELISA, Western Blot and Immunoprecipitation. Other applications not tested.
Recommended Dilutions
Immunofluorescence: 0.5-2ug/ml ELISA: 0.1-1ug/ml Western Blot: 1-3ug/ml Immunoprecipitation: 2-3ug Optimal dilutions to be determined by the researcher.
Storage and Stability
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Immunogen
Synthetic peptide derived from the internal region of the human ZO-3 protein.
Form
Supplied as a liquid in PBS, pH 7.4, 0.09% sodium azide
Purity
Purified by immunoaffinity chromatography
Specificity
Recognizes human ZO-3 protein with a molecular weight of ~130kD. Crossreactivity with other Zonula Occluden family members has not been observed. Reactivity is confirmed with human T84, HeLa, and canine MDCK cell lysates.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.

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.
USBio References
No references available
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