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AlphaLISA technology allows detecting the binding of target proteins in a highly sensitive, quantitative, reproducible and user-friendly mode. In this AlphaLISA assay, a biotinylated PDGF-BB binds to the Streptavidin-coated Alpha Donor beads, while His tagged PDGFRα is captured by Anti-His AlphaLISA Acceptor beads. When PDGF-BB binding to PDGFRα 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.
Platelet-derived growth factor (PDGF) is one of numerous growth factors that regulate cell growth and division. PDGF is a dimeric glycoprotein that can be composed of two A subunits (PDGF-AA), two B subunits (PDGF-BB), or one of each (PDGF-AB). The dimeric isoforms are differentially expressed in various cell types and their effects are mediated through two cell surface tyrosine kinase receptors, platelet-derived growth factor receptors, termed alpha (PDGFRα) and beta (PDGFRβ). Differences exist in isoform binding to each receptor. PDGF and PDGFR are involved in human cancer development and progression through autocrine stimulation of tumor cell growth. Blocking PDGF and PDGFR binding has been considered as a potential target for antitumor and antiangiogenic therapy.
|Assay Target||PDGF-BB, PDGFR|
|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.