Technical Data
Z0750-02B
ZO-1 (Tight Junction-associated Polypeptide, Zonula Occludens 1)
Description:
ZO-1 is a peripheral membrane protein that is associated with tight junctions (zonula occludentes) which are found on the cytoplasmic surface of epithelial and endothelial cell membranes. ZO-1 has a MW of 225kD and is found in two isoforms that differ by the presence or absence of an 80 amino acid region known as 'motif-a'. These two isoforms are the basis for determining two classes of tight junctions. ZO-1 is known to interact with ZO-2. ZO-1 is homologous to dlg, the Drosophila tumor suppressor. Both are members of a putative signal transduction protein family whose members contain a SH3 domain and a domain homologous to the enzyme guanylate kinase.

Applications:
Suitable for use in ELISA, Western Blot, Immunoprecipitation and Immunohistochemistry. Other applications have not been tested.

Recommended Dilutions:
Western Blot: 0.5-2ug/ml
Immunoprecipitation: 2-5ug/IP reaction
Immunohistochemistry (Paraffin/Frozen): 2-5ug/ml. To achieve appropriate staining results on formalin-fixed, paraffin-embedded tissue sections, protease pretreatment is required. Use 1-2mg/ml protease and incubate at 37°C for 10 minutes. Other enzyme and HIER pre-treatment methods may not be suitable for this antibody. Frozen sections have been successfully stained after fixation with acetone at 4°C.
Optimal dilutions to be determined by the researcher.

Positive Controls for Immunoprecipitation:
Canine MDCK and human Caco-2 cells.

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. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
TypeIsotypeCloneGrade
PabIgGAffinity Purified
SizeStorageShippingSourceHost
100ug-20°CBlue IceHumanRabbit
Concentration:
~0.25mg/ml
Immunogen:
A 69kD fusion protein corresponding to aa463-1109 of human ZO-1 cDNA. This sequence lies from the N-terminal to the 80 amino acid region (the alpha-motif) present in the a+isoform but absent in the alpha- isoform due to alternative splicing.
Purity:
Purified by immunoaffinity chromatography.
Form
Supplied as a liquid in PBS, pH 7.4, 0.1% sodium azide.
Specificity:
Recognizes human ZO-1 alpha+ and ZO-1 alpha- isoforms. Reactivity with dlg has not been characterized. No reactivity with PSD-95 or p55 has been observed on Western Blots, but potential reactivity with these proteins has not been fully characterized. Species Crossreactivtiy: mouse, rat, guinea pig, canine. Moderate reactivity with chicken.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Willott, E. et al., Am J Physiol. 262 (Cell Physiol. 31):C1119-C1124 (1992). 2. Willott, E. et al., Proc. Natl. Acad. Sci., USA 90(16):7834-7838 (1993). 3. Balda, M.S. and Anderson, J.M., Am J Physiol 264 (Cell Physiol. 33):C918-C924 (1993). 4. Tsarfaty, I, et al., Science 263:98-101 (1994). 5. Keon, BH, et al., J. Cell Biol. 134(4):1003-1018 (1996). 6. Bomsel, M. Nature Medicine 3(1):42-47 (1997). 7. Watabe-Uchida, M, et al., J. Cell Biol 142(3):847-857 (1998). 8. Hoover, KB, et al., J. Am. J. Pathol 153(6):1767-1773 (1998). 9. Rajasekaran, AK, et al., J. Cell Biol 132(3):451-463 (1996). 10. Haskins, J, et al., J. Cell Biol. 141(1):199-208 (1998). 11. Kachinsky, AM, et al., J. Cell Biol. 145:391-402 (1999). 12. Wu, S., et al., J. Proc. Natl. Acad. Sci. 95(25):14979-14984 (1998). 13. Moreira, JE, et al. J. Histochem. Cytochem. 46(7):847-854 (1998).14. Potempa, S. and Ridley, A.J. Mol Biol Cell 9:2185-2200 (1998).15. Mann, B, et al., Proc. Natl. Acad. Sci. 96:1603-1608 (1999).16. Laing, JG, et al., J. Biol. Chem. 276 (25) : 23051-23055 (2001). 17. Hirata, K., et al., J. Biol. Chem. 276 (19) : 16223-16231 (2001).18. Toyofuku, T., et al., J. Biol. Chem. 276 (3) : 1780-1788 (2001).19. Singh, U., et al., J. Biol. Chem. 275 (24): 18407-18417 (2000).20. Bazzoni, G., et al., J. Biol. Chem. 275 (27) : 20520-20526 (2000).21. Fluge, O., et al., World J. Surg. 25: 623-631 (2001).22. Hamazaki, Y., et al., J. Biol. Chem. 277 (1): 455-461 (2002).23. Duffy, H.S., et al., J. Neuroscience 20 :RC114(1-6)