Product Information

Description: Mouse Anti-Human Interferon Beta

Clone: MMHB-2

Contents: 500 μg/vial

Concentration: 0.5 mg/ml; after reconstitution with 1ml sterile PBS

Form: Lyophilized from a 0.2 µm filtered solution in phosphate-buffered saline (PBS) containing 5% trehalose

Reconstitution: Dissolve contents of the vial by injection of 1 ml sterile PBS

Antigen: human interferon beta

Isotype: Mouse IgG₁

Purification Method: Protein G affinity chromatography

Specificity: Neutralizes human interferon beta; does not neutralize human interferon alpha or gamma

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 ~7-21 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.

Due to the variation in ND₅₀ values based on cell type and assay system, we recommend each user determine the neutralizing concentration of this antibody lot in their assay system. Using an A549/EMCV (cell/virus) system, we have not verified with reasonable consistency the neutralizing concentration of this antibody (the concentration required to inhibit the antiviral effect of human interferon beta by one half).

Tested Applications: Neutralization; Direct ELISA (0.5-1.0 µg/ml); Western Blot (1-2 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 PBS, 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.

Product Information

Figure 1: Human IFN-β 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-β 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 100βl, containing antibody at the concentrations as indicated, Hu-IFN-β 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 prewarmed 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 7-21 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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