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AlphaLISA technology allows the detection of molecules of interest in buffer, cell culture media, serum and plasma in a highly sensitive, quantitative, reproducible and user-friendly mode. In an AlphaLISA assay, a Biotinylated Anti-Analyte Antibody binds to the Streptavidin-coated Alpha Donor beads, while another Anti-Analyte Antibody is conjugated to AlphaLISA Acceptor beads. In the presence of the analyte, the beads come into close proximity. The excitation of the Donor beads provokes the release of singlet oxygen molecules that triggers a cascade of energy transfer in the Acceptor beads, resulting in a sharp peak of light emission at 615 nm.
Tumor necrosis factor alpha (TNFα) is a multifunctional proinflammatory cytokine produced primarily by activated macrophages and T cells. TNFα is primarily produced as 233 Amino-Acid long TNF type II transmembrane proteins. The soluble form of TNFα can be generated from its transmembrane form via proteolytic cleavage by ADAM17. Released TNFα self-associates to form a soluble complex that exerts its function by binding the TNFR1 and TNFR2 receptors. TNFα has been observed to play a role in inflammation, apoptotic cell death, and as an important regulator of tumorigenesis and infection. TNFα dysregulation has been observed in several major diseases such as Inflammatory Bowel Disease, Depression, Cancer, and Neurological Diseases. This kit has been designed for the detection of TNFα in cynomolgus and rhesus monkey serum and cell culture supernatants.
|Assay Target Class||Cytokine|
|Experimental Type||In vitro|
|Product Brand Name||AlphaLISA|
|Shipping Condition||Blue Ice|
|Unit Size||5,000 Assay Points|
The introduction of enzyme-linked immunosorbent assays (ELISAs) in the early 1970’s offered researchers a non-radiometric immunoassay platform without compromising sensitivity. Over the last 50 years scientists have made huge strides in disease research and drug discovery and a demand for greater assay throughput and sensitivity has evolved. In response, more robust immunoassays have been developed to address some of the limitations of the standard, colorimetric ELISA.
Find out about the most common limitations of traditional ELISAs and how different ELISA alternative technologies address these limitations.
This manual explains how to run the AlphaLISA cynomolgus TNFa detection assay.