Product Information

Description: Goat Anti-Human Interferon Beta

Contents: 100 µg/vial

Concentration: 0.1 mg/ml; after reconstitution with 1ml sterile PBS.

Form: Lyophilized from a 0.2 µm filtered solution in phosphate-buffered saline (PBS).

Antigen: Recombinant human interferon beta

Isotype: Human interferon beta specific goat IgG

Purification: Human IFN beta affinity chromatography

Assay Used to Measure Bioactivity: The exact concentration of antibody required to neutralize human interferon beta activity is dependent on the cytokine concentration, cell type, growth conditions and type of activity studied. The Neutralization Dose₅₀ (ND₅₀) for this antibody is defined as that concentration of antibody required to yield one-half maximal inhibition of the cytokine activity on a responsive cell line, when that cytokine is present at a concentration just high enough to elicit a maximum response. The ND₅₀ for this antibody on human (HeLa/EMCV) cells is ~0.05- 0.2 µg/ml in the presence of 10ng/ml of human interferon beta, based on the anti-viral assay. The specific conditions are described in the figure legends.

Specificity: Neutralizes human interferon beta; does not neutralize the biological activity of human interferon alpha or gamma

Tested Applications: Neutralization; Direct ELISA (0.5-1.0 µg/ml); Western Blot (0.1-0.2 µg/ml). Optimal dilutions should be determined by each laboratory for each application.

Shipping Information

Shipping Conditions Wet ice

Physical State of Product During Shipping: Lyophilized

Special Conditions/Comments: After receipt, this product should be kept at -20°C for retention of full activity. Upon reconstitution with sterile Tris-buffered saline pH 7.3 containing 0.1% BSA, the antibody can be stored at 2-8°C for 1 month without detectable loss of activity. Reconstituted antibody can also be aliquoted and frozen at -20° to -70°C in a manual defrost freezer for 6 months without detectable loss of activity. Avoid repeated freeze-thaw cycles.

Figure 1:                                    Figure 2:

Figure 1: Human IFN-b reduces the cytopathic effect of the lytic virus EMC in a dose-dependent manner, on the human cell line, HeLa. (Meager, A. 1987, Lymphokines and Interferons, a practical approach, Clemens, M.J. Morris, A.G. and A.J.H. Gearing, eds. IRL Press, p. 129). The ED₅₀ for this effect is typically 2-5 ng/ml.

Figure 2: To measure the ability of the antibody to neutralize the bioactivity of the human interferon beta on HeLa cells, Hu-IFN-b was added to various concentrations of the antibody. The antigen-antibody mixture was added to confluent cultures of the HeLa cells in a 96 well plate. The assay mixture in a total volume of 100ml, containing antibody at the concentrations as indicated, Hu-IFN-Beta; at 10 ng/ml, was incubated at 37°C for 20-24 hours in a humidified CO₂ incubator. At the end of this incubation period, medium was aspirated from all wells and an appropriate titrated amount of the EMCV in pre-warmed culture medium was added to each test well. After another 20-24 hour incubation, the cells were fixed, stained and scored for cytopathic effect by measurement of optical densities in a microplate reader at 540 nm. The ND₅₀ of the antibody is approximately 0.5-0.2 µg/ml.

This Product is for RESEARCH USE ONLY and is not for sale or use in any commercial kit.
NOT FOR DIAGNOSTIC OR THERAPEUTIC USE, OR USE IN HUMANS.

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