Anti-Interleukin 5 (IL-5, BCDF-alpha, Eosinophil Differentiation Factor, EDF)

Interleukin 5 (IL-5) is a disulfide-linked homodimeric cytokine that is secreted by T cells. IL-5 promotes the proliferation, differentation and activation of eosinophils. It binds to a receptor complex consisting of one IL-5 specific alpha chain and one non-binding common beta chain that is shared with the receptors for GM-CSF and IL-3. Applications: Suitable for use in ELISA, Western Blot, Immunohistochemistry, and Flow Cytometry. Other applications have not been tested. Recommended Dilution: Direct ELISA: 0.5-1.0ug/ml. Detection limit foe rhIL5 is ~2ng/well., Western Blot: 1-2ug/ml. Detection limit foe rhIL5 is ~25ng/lane, Immunohistochemistry: 5-15ug/ml. Human peripheral blood lymphocytes or activated T-cells may be fixed for 20 minutes at RT with freshly prepared 1-2% paraformaldehyde/PBS (pH 7.4). Incubate cells for 60 minutes at RT with I8427-03E. , Flow Cytometry: 25ug/mL. For Flow Cytometry on PMA plus Ca2+ ionomycin-treated PBMCs, cells must first be fixed and permeabilized using 4% paraformaldehyde and 0.1% saponin in PBS to detect intracellular staining of IL-5. Dilute I8427-03E to 25ug/mL and add 10ul of the diluted solution to 1-5x10e5 cells in a total reaction volume not exceeding 200ul. Visualize staining by adding a secondary developing reagent such as anti-mouse IgG conjugated to a fluorochrome*. , Optimal dilutions to be determined by the researcher. Recommended Secondary Antibodies: I1904-65V IgG, F(ab')2 (FITC) Pab RbxMo, I1904-66A IgG, F(ab')2, X-Adsorbed (FITC) ) Pab RbxMo, , Storage and Stability:, Lyophilized powder may be stored at -20°C. Stable for 12 months at -20°C. Reconstitute by adding sterile 40-50% glycerol, PBS. Aliquot and store at -20°C. Reconstituted product is stable for 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.,