Materials Provided:

• Pre-coated microtiter plate(s)
• Plate Sealers
• Wash Solution Concentrate
• Human IFN-Beta Standard, 100,000 pg/ml
• Sample Diluent 
• Antibody Concentrate 
• HRP Conjugate Concentrate 
• Concentrate Diluent
• TMB Substrate Solution
• Stop Solution

Additional Materials Required (Not Provided)

• Microplate reader capable of reading an OD at a wavelength of 450 nm
• Variable volume microtiter pipettes
• Adjustable multichannel pipette (50-300 μl)
• Reagent reservoirs
• Wash bottle or plate washing system
• Distilled or deionized water
• Serological pipettes (1, 5, 10 or 25 ml)
• Disposable pipette tips (polypropylene)
• Plate shaker

Background

Following secretion, Interferon Beta binds to type I interferon receptors on proximal or distal cells activating the JAK1-STAT signaling pathway. Activation of this signal transduction pathway leads to the expression of 2’-5’ oligoadenylate synthetases (2’5’ OAS), protein kinase R (PKR), MxA proteins, and interferon regulatory factor 7 (IRF-7). The upregulation of IRF-7 expression can exert positive feedback on IFN-β production, whereas the induction of 2’5’ OAS activates a latent endonuclease known as RNase L. RNase L cleaves both viral and cellular single-stranded mRNA, thereby limiting viral replication and dissemination.

IFN-beta is present as a single gene in humans. The biological activity of IFN-beta can be functionally similar when compared to other type I IFNs, but can also exhibit distinct gene expression patterns as compared to IFN-alpha. In vitro binding studies suggest that IFN-beta binds to the Type I IFN receptor with higher affinity than the other Type I IFNs and this may regulate receptor internalization in a different manner. IFN-β has long been known to inhibit viral replication as part of the body’s innate antiviral response and it is currently used as a therapeutic for treatment of relapsing-remittent multiple sclerosis and some tumors.

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Background Literature:

1. Staehelin et al. (1981) Methods in Enzymology, Vol. 79 (Pestka, ed.), Academic Press, New York, 589-595.
2. Kelder et al. (1986) Methods in Enzymology, Vol. 119 (Pestka, ed.), Academic Press, New York, 582-587.
3. Human IFN-β international reference standard provided by the NIH, reference no. Gb23-902-531. Pestka (1986) Methods in Enzymology, Vol. 119 (Pestka, ed.), Academic Press, New York, 14-23.
4. Moschera et al. (1981) Methods of Enzymology, Vol. 119 (Pestka, ed.), Academic Press, New York, 177-183.