Cynomolgus/Rhesus IFN-Alpha ELISA Kit (Serum, Plasma, TCM)

Materials Provided

• Pre-coated microtiter plate
• Plate Sealers
• Wash Solution Concentrate
• Rhesus/Cyno IFN-Alpha 2 Standard, 10,000 pg/ml
• Standard Diluent
• Sample 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

Application Note

• VeriKine Cynomolgus/Rhesus IFN-α Serum ELISA Performance Evaluation

White Paper (by request)

• Utility and Application of the VeriKine Cynomolgus/Rhesus Interferon Alpha Serum ELISA

Background

Interferons (IFNs) are a group of cytokines which exhibit pleiotropic activities that play major roles in both innate and adaptive immunity. Type I IFNs consist of at least one IFN-β gene and protein as well as multiple IFN-α genes and proteins in most vertebrate species¹.

IFN-α expression and secretion are primarily induced by signaling events processed through pattern recognition receptors such as the Toll-like and RIG-I like receptors (TLR and RLR, respectively). While IFN-α can be produced by most cell types, strong evidence suggests that plasmacytoid dendritic cells are a major source of IFN-α in vivo².

The interferon alpha gene family is comprised of multiple distinct genes that occupy a single chromosomal locus. All sequenced vertebrate species possess a large number of distinct alpha genes suggesting a biological significance to maintaining multiple copies of IFN-Alpha genes. Published reports have shown the expression patterns of the individual IFN-Alpha proteins and their cell-specific function can vary suggesting unique properties of each gene in a cell-dependent manner.

Following expression and secretion, IFN-α binds to a heterodimeric receptor chain consisting of IFNAR1 and IFNAR2 subunits on proximal and distal cell surfaces. Receptor binding promotes a signal transduction cascade consisting of components of the JAK-STAT signaling pathway. Hundreds of genes are regulated subsequent to binding of the IFNAR receptor subunits to IFN-α, thus leading to the antiviral, anti-proliferative and immunomodulatory activities of the cytokine.

Two nonhuman primate species, Rhesus (Macaca mulatta) and Cynomolgus (Macaca fascicularis) macaques, are sufficiently genetically similar to humans that they are emerging as highly relevant animal models for studying various aspects of human physiology³. For example, macaques provide valuable surrogate models for examining the pathogenicity of human viral infections such as the 1918 pandemic influenza virus⁴. Furthermore, macaques are included in the evaluation of many new therapeutic agents aimed at modulating host immunity to either enhance or dampen immune responses^5,6.

These primates also serve as important immunotoxicological models in the testing of human pharmaceuticals⁷.