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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.
Lymphocyte-activation gene 3 (LAG-3), also known as CD223, is a cell surface protein that affects T-cell function in many ways. LAG-3 is expressed on natural killer cells, plasmacytoid dendritic cells, B-cells, and activated T-cells. LAG-3 works in similar fashion to CTLA-4 and PD-1 in that it negatively regulates cellular proliferation and Treg suppression. Maturation and activation of dendritic cells is also known to be affected by LAG-3 expression. Soluble forms are in clinical development to activate dendritic cells. Anti-LAG-3 antibodies are undergoing studies for anti-cancer and autoimmune response.
|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.
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:
Various cytokines are secreted during an active immune response that can have modulatory effects on target cell populations, including interferon gamma (IFN-ɣ), tumor necrosis factor alpha (TNFa) and several interleukins.
In this application note, you will learn how we investigated:
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 LAG-3 detection assay.