Technical Data
Suitable for use in ELISA, Western Blot, Immunofluorescence, Immunoprecipitation and Immunohistochemistry. Other applications not tested.

Recommended Dilutions:
ELISA: 0.1-1ug/ml
Western Blot: 1-3ug/ml
Immunofluorescence: 4-20ug/ml
Immunoprecipitation: ~5ug/IP reaction
Immunohistochemistry (FFPE): 2-5ug/ml; Protease pretreatment of sections is required prior to staining. Dilute protease to 1-2mg/ml and incubate at 37°C for 10 minutes. Other enzyme and HIER pretreatment methods may not be suitable for this antibody.
Optimal dilutions to be determined by the researcher.

Recommended IF fixation and staining protocol:
1.“Pre-extract” the cells on ice for 2 min with an ice-cold buffer containing 0.2% Triton X-100.
2. Fix cells in 3% paraformaldehyde in PBS for 30 min at RT.
3. Permeabilize sample with 0.05% Triton X-100 in PBS for 5 min on ice.
4. Incubate samples with the desired concentration of O5204 (4-20ug/ml) on ice for 1 hour.
5. Incubate with IgG (FITC) Pab Gt x Rb for 30 min at RT.

Recommended Secondary Antibodies:
I1904-40D: IgG, H&L (FITC) Pab Gt xRb
I1904-41: IgG, X-Adsorbed (FITC) Pab Gt xRb
I1904-46N: IgG, H&L (FITC) Pab Gt xRb

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.
PabIgGAffinity Purified
100ug-20°CBlue IceHumanRabbit
Fusion protein containing the C-terminal 150aa of human occludin.
Purified by immunoaffinity chromatography.
Supplied as a liquid in PBS, pH 7.4, 0.1% sodium azide.
Recognizes human occludin at ~65kD. Reactivity of this antibody with the occludin protein has been confirmed by Western Blot using total cell lysates derived from MDCK cells, human T84 cells, human Caco-2 and rat fibroblasts transfected with a plasmid encoding human occludin. Species Crossreactivity: mammalian, mouse, rat and canine
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. Anderson, J.M., et al., Curr. Opin. Cell. Biol. 5: 772-778 (1993). 2. Gumbiner, B., Am. J. Physiol. 123: 1631-1633 (1993). 3. Farquhar, M. et al, Cell. Biol. 17: 375-409 (1963). 4. Stevenson, B.R., et al., J. Cell. Biol. 103: 755-766 (1986). 5. Anderson, J.M., et al., J. Cell. Biol. 106: 141-1149 (1988). 6. Itoh, M., et al., J. Cell. Biol. 115: 1449-1462 (1993). 7. Gumbiner, B., et al., Proc. Natl. Acad. Sci. USA 88: 3460-3464 (1991). 8. Citi, S., et al., Nature 33: 272-276 (1988). 9. Zhong, Y., et al., J. Cell. Biol. 120: 477-483 (1993). 10. Keon, B.H., et al., J. Cell. Biol. 134: 1003-1018 (1996). 11. Furuse, M., et al., J. Cell Biol. 123: 1777-1788 (1993). 12. Furuse, M., et al., J. Cell. Biol. 127: 1617-1626 (1994). 13. Ando-Akatsuka, Y., et al., J. Cell. Biol. 133: 43-47 (1996). 14. Balda, et al., J. Cell Biol. 134(4): 1031-1049 (1996). 15. Jou, T.S. and Nelson W.J., J. Cell Biol. 14(1): 101-115 (1998). 16. Martin-Padura, I., et al., J. Cell Biol. 142(10): 117-127 (1998). 17. Fanning, A.S., et al., J. Biol. Chem. 275(45): 29,745-29,753 (1998). 18. Potempa, S., et al., Mol. Biol. Cell 9(8): 2185-2200 (1998). 19. Sakakibaro, A., et al., J. Cell Biol. 137(6): 1393-1401 (1997). 20. Wu, S., et al., Proc. Natl. Acad. Sci. 95(25): 14,979-14,984 (1998).).