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

Immunohistochemistry (FFPE): 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.

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.

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

Store product at 4°C if to be used immediately within two weeks. For long-term storage, aliquot to avoid repeated freezing and thawing and store at -20°C. Aliquots are stable at -20°C for 12 months after receipt. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. Caution: FITC conjugates are sensitive to light. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Note: Applications are based on unconjugated antibody.

Fusion protein containing the C-terminal 150aa of human occludin.

Supplied as a liquid in PBS, pH 7.2. No preservatives added. Labeled with Fluorescein isothiocyanate (FITC).

Purified by immunoaffinity chromatography.

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