| Catalog # | O5204 |
| Applications | 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. |
| CAS Number | n/a |
| Clone Type | Polyclonal |
| Isotype | IgG |
| Host | Rabbit |
| Source | Human |
| Concentration | ~0.25mg/ml |
| Form | Supplied as a liquid in PBS, pH 7.4, 0.1% sodium azide. |
| Purity | Purified by immunoaffinity chromatography. |
| Immunogen | Fusion protein containing the C-terminal 150aa of human occludin. |
| Specificity | 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 |
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| Important Note | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
