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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||100 Assay Points|
One approach to immunotherapy is the modulation of immune checkpoints that are critical in regulating the degree and duration of immune system responses and preventing autoimmunity.
In this application note, you will learn:
Immune checkpoints serve a critical role in the immune system to prevent autoimmunity and manage the degree and duration of an immune response. Cytotoxic T-Lymphocyte-associated protein 4 (CTLA-4 or CD152) is an inhibitory transmembrane protein involved in an immune checkpoint of significant interest for therapeutic development. When CTLA-4 is expressed and competes with CD28, the immune system response is downregulated. As a result of this immune system response balance, immune checkpoints provide an opportunity for therapeutic intervention to modulate immune system activity.
There is a high demand for new drugs to block CTLA-4 and modulate immune system activity. In this application note, we demonstrate how to screen for novel CTLA-4 blocking drugs by utilizing the AlphaLISA CTLA-4/CD80 binding assay.
This manual describes how to run an AlphaLISA human HVEM detection assay.