Materials Provided

• Pre-coated microtiter plate(s)
• Plate Sealers
• Wash Solution Concentrate
• Human IFN-Alpha Standard, 10,000 pg/ml
• Dilution Buffer
• Antibody Concentrate
• HRP Conjugate Concentrate
• Concentrate Diluent
• TMB Substrate
• 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

Tips, Tools and Troubleshooting:

• Human IFN-α ELISA Subtype Specificities Comparison Chart

Background

Interferons (IFNs) are a family of mammalian cytokines initially characterized by their ability to inhibit viral infection. They are synthesized and secreted by most cell types in response to pathogens. In addition to their antiviral properties, IFNs have also been shown to exhibit anti-proliferative, immunomodulatory, and many other activities.

In humans, IFN-α consists of a group of proteins that are greater than 85% homologous by amino acid sequence. Numerous individual human IFN-α subtypes have been identified; many display different properties. It remains unclear why there are multiple IFN-α subtypes. A variety of studies suggested they possess overlapping but also unique sets of biological activities.

35 Citations

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12. Dowell, Nicholas, et al. (2016). Acute Changes in Striatal Microstructure Predict the Development of Interferon-Alpha Induced Fatigue. Biological Psychiatry, 9 pgs. PMID: 26169252. (link)


13. Huang, Cheng, et al. (2015). Highly Pathogenic New World and Old World Human Arenaviruses Induce Distinct Interferon Responses in Human Cells. JVI, 10 pgs. PMID: 25926656. (link)


14. Bonaparte, Sarah, et al. (2015). HPV-Positive Cervical Cancer Cells Show Generally Reduced Levels of IFN-Alpha. Belmont University, 11 pgs. PMID: no PMID. (link)


15. Huang, Yanyan, et al. (2014). Hcmv-miR-UL112 Attenuates NK Cell Activity by Inhibition Type I Interferon Secretion. Immunology Letters, 30 pgs. PMID: 25530545. (link)


16. Lederle, Alexandre, et al. (2014). Neutralizing Antibodies Inhibit HIV-1 Infection of Plasmacytoid Dendritic Cells by an Fc-gamma-Rlla Independent Mechanism and Do Not Diminish Cytokines Production. Scientific Reports, 10 pgs. PMID: 25132382. (link)


17. Vogelsang, Petra, et al. (2014). Altered Phenotype and Stat1 Expression in Toll-Like Receptor 7/8 Stimulated Monocyte-Derived Dendritic Cells from Patients with Primary Sjoergen's Syndrome. Arthritis Research & Therapy, 22 pgs. PMID: 25113744. (link)


18. Maeyama, Jun-ichi, et al. (2014). A Palindromic CpG-Containing Phosphodiester Oligodeoxynucleotide as a Mucosal Adjuvant Stimulates Plasmacytoid Dendritic Cell-Mediated T_H1 Immunity. PLOS One, 10 pgs. PMID: 24586411. (link)


19. Stone, Amy, et al. (2013). Hepatitis C Virus Pathogen Associated with Molecular Pattern (PAMP) Triggers Production of Lambda-Interferons by Human Plasmacytoid Dendritic Cells. PLOS One, 13 pgs. PMID: 23637605. (link)


20. Chen, Qian, et al. (2013). Interleukin-27 is a potent inhibitor of cis HIV-1 replication in monocyte-derived dendritic cells via a type I interferon-independent pathway. PLOS One, 10 pgs. PMID: 23527130. (link)


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26. Xu, Yongfen, et al. (2009). HBsAg inhibits TLR9-mediated activation and IFN-alpha production in plasmacytoid dendritic cells. Molecular Immunology, 7 pgs. PMID: (link)


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32. Abel, Kristina, et al. (2005). Deoxycytidyl-Deoxyguanosine Oligonucleotide Classes A, B, and C Induce Distinct Cytokine Gene Expression Patterns in Rhesus Monkey Peripheral Blood Mononuclear Cells and Distinct Alpha Interferon Responses in TLR9-Expressing Rhesus Monkey Plasmacytoid Dendritic Cells. Clinical and Diagnostic Laboratory Immunology, 16 pgs. PMID: 15879022. (link)


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Background Literature:

1. Staehelin, T., Stähli, C., Hobbs, D.S., and Pestka, S. (1981) “A Rapid Quantitative Assay of High Sensitivity for Human Leukocyte Interferon with Monoclonal Antibodies,” in Methods in Enzymology, Vol. 79 (S. Pestka, ed.), Academic Press, New York, 589-595.


2. Kelder, B., Rashidbaigi, A., and Pestka, S. (1986) “A Sandwich Radioimmunoassay for Human IFNa” in Methods in Enzymology, Vol. 119 (S. Pestka, ed.), Academic Press, New York, 582-587.


3. Human IFN-α international reference standard provided by the NIH, reference no. Gxa01-901-535. Pestka, S. (1986) “Interferon Standards and General Abbreviations,” in Methods in Enzymology, Vol. 119 (S. Pestka, ed.), Academic Press, New York, 14-23.


4. Rubinstein, M., Levy, W.P., Moschera, J.A., Lai, C.-Y., Hershberg, R.D., Bartlett, R.T., and Pestka, S. (1981) “Human Leukocyte Interferon: Isolation and Characterization of Several Molecular Forms,” Arch. Biochem. Biophys. 210, 307-318.


5. Hobbs, D.S. and Pestka, S. (1982) “Purification and Characterization of Interferons from a Continuous Myeloblastic Cell Line,” J. Biol. Chem. 257, 4071-4076.