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, 0.09% 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.
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