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Human IFN-Alpha ELISA Kit (TCM)

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Human IFN-Alpha ELISA Kit (TCM)

Catalog Number: 41100

This often-cited ELISA assay quantifies human IFN-Alpha in tissue culture media (TCM) from 12.5-500 pg/ml or from an extended range of 156-5000 pg/ml.

 

Click here to see which PBL Human IFN-Alpha ELISA kit is best fit for your research need.

 

Product Name: VeriKine Human Interferon-Alpha ELISA Kit (TCM)

$585.00

Pack Size
Product Info

Matrix Compatibility Tissue Culture Media (TCM)
Assay Range 12.5 - 500 pg/ml or 156 - 5000 pg/ml (Extended Range)
Assay Length 3 hours, 15 minutes
Specificity Human Interferon Alpha

 

The VeriKine Human IFN-Alpha ELISA Kit has been developed to quantify levels of human IFN-alpha in cell or tissue culture media (TCM) using a sandwich enzyme-linked immunoassay (ELISA). The kit is based on an ELISA with anti-detection antibody conjugated to horseradish peroxidase (HRP). Tetramethyl-benzidine (TMB) is the substrate. The assay is based on the international reference standard for human IFN-Alpha provided by the National Institutes of Health.

 

*For global subtype detection and additional sensitivity, we recommend our Human IFN-Alpha All-Subtype TCM ELISA Kit, High Sensitivity (Cat. No. 41135-1).

 

*For global subtype detection, additional sensitivity, and complex biological matrices such as plasma and serum, we recommend our Human IFN-Alpha All-Subtype ELISA Kit, High Sensitivity (Cat. No. 41115-1).

Specifications

CVs
Inter-Assay ≤ 8%
 
Intra-Assay ≤ 8%
Cross-reactivity

No cross-reactivity against

  • Human IFN-Gamma, IFN-Beta or IFN-Omega
  • Mouse or Rat IFN-Alpha, IFN-Beta or IFN-Gamma
  • Bovine IFN-Tau
Synonyms IFN alpha, IFN-a, interferon alpha, IFN alpha 2, IFN-a2, interferon alpha 2, Leukocyte interferon, Leukocyte IFN, alpha interferon, interferon alfa, IFN alfa, interferon alpha 2a, IFN alpha 2a, interferon a, interferon alpha 2b, IFN alpha 2b, Type I interferon alpha, Type I IFN alpha
Storage 2-8°C
Expiration Date Twenty-one months from the date of manufacture
Shipping Condition Wet Ice

 

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

Tech Info & Data

Tips, Tools and Troubleshooting:

 

Background:

 

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

 

When IFN interacts with its cognate receptor, a signal is rapidly transmitted within the cell, often producing an antiviral state. The primary signal transduction cascade promoted by type I IFNs is the JAK1-STAT pathway.

 

Activation of this signal transduction pathway leads to increased gene expression including (2’-5’) oligoadenylate synthetases, Mx proteins, and protein kinase R (PKR) that protect the cell from viral infection.

Citations

57 Citations

 

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  2. Zhang, Z., et al., (2021), "IRF7 and RNH1 are modifying factors of HIV-1 reservoirs: a genome-wide association analysis", BMC Medicine, 19:282, DOI:10.1186/s12916-021-02156-5. (link)

  3. Sabree, SA., et al., (2021), "Direct and indirect immune effecrs of CMP-001, a virus-like particle containing a TLR-9 agnoist", Journal of ImmunoTherapy of Cancer, 9:e002484, DOI: 10.1136/jitc-2021-002484 (link)

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  6. Loughran, Sinead, et al. (2018). Influenza infection directly alters innate IL-23 and IL-12p70 and subsequent IL-17A and IFN-γ responses to pneumococcus in vitro in human monocytes. PLOS One, 17 pgs. PMID: 30192848. (link)

  7. Moskwa, Sylwia, et al. (2018). Innate Immune Response to Viral Infections in Primary Bronchial Epithelial Cells is Modified by the Atopic Status of Asthmatic Patients. AAIR, 11 pgs. PMID: 29411555. (link)

  8. Svensson, Alexandra, et al. (2017). Maturation-Dependent Expression of AIM2 in Human B-Cells. PLOS One, 13 pgs. PMID: 28809949. (link)

  9. Wang, et al. (2017). Intracellular DNA sensing pathway of cGAS-cGAMP is decreased in human newborns and young children. Molecular Immunology. PMID: 28412547. (link)

  10. Schuh, Elisabeth, et al. (2017). Human Plasmacytoid Dendritic Cells Display and Shed B Cell Maturation Antigen upon TLR Engagement. Journal of Immunology, 9 pgs. PMID: 28283566. (link)

  11. Herawati, Neng, et al. (2017). Overproduction, Characterization, and Preliminary Antiproliferative Activity Determination of Non-Tagged Recombinant Human Interferon Alpha-2a Produced in Pichia pastoris. Nusantara Bioscience, 5 pgs. PMID: no PMID. (link)

  12. Abd-Elraheem, Sabah, et al. (2017). Estimation of Serum Interferon Alpha and Toll-Like Receptors 3 (TLRs 3) Gene Expression in Egyptian Patients with Liver Cirrhosis. JBC, 8 pgs. PMID: no PMID. (link)

  13. Hammerstad, Sara, et al. (2016). Hepatitis C Virus E2 Protein Induces Upregulation of Il-8 Pathways and Production of Heat Shock Proteins in Human Thyroid Cells. Endocrine Society, 18 pgs. PMID: 27860532. (link)

  14. Lee, Joo Youn, et al. (2016). Circulating Exosomes from Patients with Systemic Lupus Erythematosus Induce a Proinflammatory Immune Response. Arthritis Research & Therapy, 8 pgs. PMID: 27852323. (link)

  15. Bam, Rujuta, et al. (2016). TLR7 Agonist GS-9620 Is a Potent Inhibitor of Acute HIV-1 Infection in Human Peripheral Blood Mononuclear Cells. Antimicrobial Agents and Chemotherapy, 14 pgs. PMID: 27799218. (link)

  16. Millrine, David, et al. (2016). Rabex-5 is a lenalidomide target molecule that negatively regulates TLR-induced type 1 IFN production. PNAS, 6 pgs. PMID: 27601648. (link)

  17. Jang, Mihue, et al. (2016). A RNA Nanotechnology Platform for a Simultaneous Two-in-One siRNA Delivery and its Application in Synergistic RNAi Therapy. Scientific Reports, 16 pgs. PMID: 27562435. (link)

  18. Nantachit, Nattika, et al. (2016). Responses of Primary Human Nasal Epithelial Cells to EDIII-DENV Stimulation: The First Step to Intranasal Dengue Vaccination. Virology Journal, 8 pgs. PMID: 27538960. (link)

  19. Perez-Pinera, Pablo, et al. (2016). Synthetic Biology and Microbioreactor Platforms for Programmable Production of Biologics at the Point-of-Care. Nature Communications, 10 pgs. PMID: 27470089. (link)

  20. Goodier, Martin, et al. (2016). Influenza Vaccination Generates Cytokine-Induced Memory-Like NK Cells: Impact of Human Cytomegalvirus Infection. Journal of Immunology, 14 pgs. PMID: 27233958. (link)

  21. Chen, Qing, et al. (2016). Carcinoma-astrocyte gap junctions promote brain metastasis by cGAMP transfer. Nature. PMID: 27225120. (link)

  22. Borhis, Gwenoline, et al. (2016). Plasmacytoid dendritic cells and myeloid cells differently contribute to B-cell-activating factor belonging to the tumor necrosis factor superfamily overexpression during primary HIV infection. Wolters Kluwer.  PMID: 26558721. (link)

  23. Deymier, Martin, et al. (2015). Heterosexual Transmission of Subtype C HIV-1 Selects Consensus-Like Variants without Increased Replicative Capacity or Interferon-α Resistance. PLOS Pathogens, 22 pgs. PMID: 26378795. (link)

  24. Jang, Mihue, et al. (2015). Design of a Platform Technology for Systemic Delivery of siRNA to Tumours Using Rolling Circle Transcription. Nature Communications, 12 pgs. PMID: 26246279. (link)

  25. Han, Tao, et al. (2015). Set7 facilitates hepatitis C virus replication via enzymatic activity-dependent attenuation of the IFN-related pathway. Journal of Immunology, 13 pgs. PMID: 25681344. (link)

  26. Wu, Zeguang (2015). Natural killer cells can inhibit the transmission of human cytomegalovirus in cell culture by using mechanisms from innate and adaptive immune responses. JVI, 12 pgs. PMID: 25540381. (link)

  27. Wolstein, Orit, et al. (2014). Preclinical Safety and Efficacy of an Anti-HIV-1 Lentiviral Vector Containing a Short Hairpin RNA to CCR5 and the C46 Fusion Inhibitor. The American Society of Gene & Cell Therapy, 14 pgs. PMID: 26015947. (link)

  28. Hsu, Yu-Lin, et al. (2014). Up-Regulation of Galectin-9 Induces Cell Migration in Human Dendritic Cells Infected with Dengue Virus. Journal of Cellular and Molecular Medicine, 12 pgs. PMID: 25754930. (link)

  29. Drewes, Julia, et al. (2014). Attenuation of Pathogenic Immune Responses During Infection with Human and Simian Immunodeficiency Virus (HIV/SIV) by the Tetracycline Derivative Minocycline. PLOS One, 15 pgs. PMID: 24732038. (link)

  30. Cai, Xinming, et al. (2014). PIKfyve, a class III lipid kinase, is required for TLR-induced type I IFN production via modulation of ATF3. Journal of Immunology, 8 pgs. PMID: 24600036. (link)

  31. Benedict, et al. (2014). Use of monitoring levels of soluble forms of cytokeratin 18 in the urine of patients with superficial bladder cancer following intravesical Ad-IFNα/Syn3 treatment in a phase l study. Nature, 4 pgs. PMID: 24503570. (link)

  32. Leng, Qixin, et al. (2012). Buffering capacity and size of siRNA polyplexes influence cytokine levels. American Society of Gene & Cell Therapy, 9 pgs. PMID: 23032972. (link)

  33. Shukla, Nikunj M, et al. (2012). Potent adjuvanticity of a pure TLR7-agonistic imidazoquinoline dendrimer. PLOS One, 11 pgs. PMID: 22952720. (link)

  34. Osterlund, Pamela, et al. (2012). Incoming influenza A virus evades early host recognition, while influenza B virus induces interferon expression directly upon entry. JVI, 11 pgs. PMID: 22855501. (link)

  35. Hayes, Melissa, et al. (2012). Pathogenic Old World Arenaviruses Inhibit TLR2/Mal-Dependent Proinflammatory Cytokines In Vitro. JVI, 12 pgs. PMID: 22532679. (link)

  36. Haseley, et al. (2012). Extracellular Matrix Protein CCN1 Limits Oncolytic Efficacy in Glioma. Cancer Research, 9 pgs. PMID: 22282654. (link)

  37. Zhu, Rui-Yu, et al. (2011). Quantitative detection of fusion protein rIFN-β-HSA by a sandwich ELISA. African Journal of Biotechnology, 7 pgs. PMID: no PMID.

  38. Dzopalic, Tanja, et al. (2010). Loxoribine, a selective Toll-like receptor 7 agonist, induces maturation of human monocyte-derived dendritic cells and stimulates their Th-1- and Th-17-polarizing capability. International Imunopharmacology, 6 pgs. PMID: 20817120. (link)

  39. Tassone, Laura, et al. (2010). Defect of Plasmacytoid Dendritic Cells in Warts, Hypogammaglobulinemia, Infections, Myelokathexis (WHIM) Syndrome Patients. Blood Journal, 5 pgs. PMID: 20736454. (link)

  40. Golding, Amit, et al. (2010). Interferon-alpha Regulates the Dynamic Balance Between Human Activated Regulatory and Effector T Cells: Implications for Antiviral and Autoimmune Responses. Immunology, 11 pgs. PMID: 20465564. (link)

  41. Liang, Xueqing, et al (2010). Toll-like receptor 9 signaling by CpG-B oligodeoxynucleotides induces an apoptotic pathway in human chronic lymphocytic leukemia B cells.. Blood Journal, pgs. PMID: 20339095. (link)

  42. Takahashi, Ken, et al. (2010). Plasmacytoid dendritic cells sense hepatitis C virus–infected cells, produce interferon, and inhibit infection. PNAS, 6 pgs. PMID: 20231459. (link)

  43. Chavez, Deborah, et al. (2009). Antiviral activity and host gene induction by tamarin and marmoset interferon-α and interferon-γ in the GBV-B primary hepatocyte culture model. Virology, 11 pgs. PMID: 19501869. (link)

  44. Chase, Jennifer, et al. (2009). Direct and indirect impairment of human dendritic cell function by virulent Francisella tularensis Schu S4. Infection and Immunity, 16 pgs. PMID: 18981246. (link)

  45. Calcedo, Roberto, et al. (2008). Host Immune Responses to Chronic Adenovirus Infections in Human and Nonhuman Primates. JVI, 10 pgs. PMID: 19116257. (link)

  46. Hooks, John, et al. (2008). IFN-beta provides immuno-protection in the retina by inhibiting ICAM-1 and CXCL9 in retinal pigment epithelial cells. Journal of Immunology, 9 pgs. PMID: 18322185. (link)

  47. Xu, Jianfeng, et al. (2007). High-yields and extended serum half-life of human interferon alpha2b expressed in tobacco cells as arabinogalactan-protein fusions. Biotechnology and Bioengineering, 13 pgs. PMID: 17328066. (link)

  48. Xu, Dongsheng, et al. (2006). The Latent Membrane Protein 1 of Epstein-Barr Virus (EBV) Primes EBV Latency Cells for Type I Interferon Production. JBC, 7 pgs. PMID: 16469740. (link)

  49. Francois, Catherine, et al. (2005). Quantification of Different Human Alpha Interferon Subtypes and Pegylated Interferon Activities by Measuring MxA Promoter Activation. Antimicrobial Agents and Chemotherapy, 6 pgs. PMID: (link)

  50. Kuchtey, John, et al. (2005). Enhancement of dendritic cell antigen cross-presentation by CpG DNA involves type I IFN and stabilization of class I MHC mRNA. Journal of Immunology, 11 pgs. PMID: 16081792. (link)

  51. Hartmann, Gunther, et al. (2005). CpG oligonucleotides induce strong humoral but only weak CD4+ T cell responses to protein antigens in rhesus macaques in vivo. Vaccine, 7 pgs. PMID: 15837237. (link)

  52. Schmidt, Kerstin N, et al. (2004). APC-independent activation of NK cells by the Toll-like receptor 3 agonist double-stranded RNA. Journal of Immunology, 7 pgs. PMID: 14688319. (link)

  53. Qu, Chunfeng, et al. (2003). Autocrine type I IFN and contact with endothelium promote the presentation of influenza A virus by monocyte-derived APC. Journal of Immunology, 10 pgs. PMID: 12517968. (link)

  54. Osborn, Blaire, et al. (2002). Pharmacokinetic and pharmacodynamic studies of a human serum albumin-interferon-alpha fusion protein in cynomolgus monkeys. The Journal of Pharmacology and Experimental Therapeutics, 9 pgs. PMID: 12388634. (link)

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  56. Jabs, William, et al. (1999). Failure to detect antiviral activity in serum and plasma of healthy individuals displaying high activity in ELISA for IFN-alpha and IFN-beta. Journal of Interferon & Cytokine Research, 7 pgs. PMID: no PMID.

 

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

Documentation

Protocol, Certificate of Analysis (CoA), Safety Data Sheet (SDS)
41100 Protocol (Full)

41100 Protocol (Full)

41100-1 CoA

41100-1 Certificate of Analysis (CoA)

41100-2 CoA

41100-2 Certificate of Analysis (CoA)

41100 SDS

41100 Safety Data Sheet