The AlphaLISA® CTLA-4/CD86 binding kit is designed for the detection of binding activity between human CTLA-4 and CD86, using a fast and simple homogeneous AlphaLISA assay (no wash steps). This assay can be used to screen for small molecules that inhibit binding, as a competitive ligand binding (CLB) assay to screen therapeutic blocking antibodies, and for potency assays.
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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 CD86 binds to the Streptavidin-coated Alpha Donor beads, while His tagged CTLA-4 is captured by Anti-His AlphaLISA Acceptor beads. When CD86 binding to CTLA-4 happens, Donor beads and Acceptor 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.
Human Cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), also known as CD152 (cluster of differentiation 152), is a cell membrane receptor and a member of immunoglobulin superfamily. CTLA-4 is expressed once a T cell becomes active and modulates T cell signals by blocking the CD80 (B7.1) and CD86 (B7.2) ligands from binding to CD28. CTLA-4, functioning as an immune checkpoint, downregulates T cell immune responses. Because of its profound inhibitory role blocking CTLA-4 and CD80 or CD86 binding has been considered as promising therapeutic target for human autoimmune disease and cancers.
|Assay Target Class||Protein|
|Product Brand Name||AlphaLISA|
|Shipping Condition||Blue Ice|
|Unit Size||5,000 Assay Points|
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.