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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.
Herpesvirus entry mediator (HVEM), also known as tumor necrosis factor receptor (TNFR) superfamily 14 and CD270, is a cell surface membrane protein that regulates T-cell immune responses by activating both inhibitory and inflammatory signaling pathways. HVEM is widely expressed on the plasma membranes of hematopoietic lineage cells, such as natural killer cells, monocytes, dendritic cells, neutrophils, T-cells and B cells. This membrane protein functions as a receptor and or ligand for BTLA, CD160, LIGHT and lymphotoxcin-alpha. Although little is known of the correlation of HVEM and cancer, it has been proven that elevated levels of HVEM have been associated in patients with autoimmune and allergic diseases such as atopic dermatitis, rheumatoid arthritis, and allergic asthma. This kit is designated to detect and quantify the levels of HVEM in cell culture supernatant and serum.
|Assay Target Class||Protein|
|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 describes how to run an AlphaLISA human HVEM detection assay.