The AlphaLISA® human TNFα/TNFR2 binding kit is designed for the detection of binding activity between human TNFα and TNFR2, 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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The AlphaLISA® human TNFα/TNFR2 binding assay uses anti-6xHis AlphaLISA Acceptor beads to capture the His-tagged TNFR2 and Streptavidin-coated donor beads to capture the biotinylated TNFα. Donor beads and Acceptor beads come into proximity through TNFα binding to TNFR2. Excitation of the Donor beads provokes the release of singlet oxygen that triggers a cascade of energy transfer reactions in the Acceptor beads, resulting in a sharp peak of light emission at 615 nm.
Tumor necrosis factor alpha (TNFα) is an immunity-modulating cytokine that is required for defense against infectious diseases and carcinogenesis. TNFα activates signals through its two receptors [TNF receptor 1 (TNFRI) and TNF receptor 2 (TNFRII)]. TNFRI is ubiquitously expressed and can be fully activated by both the membrane-bound and soluble trimeric forms of TNFα, whereas TNFRII is found mostly on certain populations of immune cells and respond to the membrane-bound form of the TNFα homotrimer. However, a block of TNFα-mediated host defense often increases the risk of bacterial or viral infection or of the development of lymphoma. Thus, a thorough understanding of the function of the TNFα-TNFR binding is important for the design of optimal therapies against the various TNFα-related autoimmune diseases.
|Assay Target Class||Cytokine, Protein|
|Product Brand Name||AlphaLISA|
|Unit Size||500 Assay Points|
Mouse pharmacological models continue to play a large role in the study of human disease, and mouse tool reagents have shown high utility in immunology and cancer research for decades. It can often be quicker to learn about immunology and the regulation of immune responses using a syngeneic mouse model. However, working in mouse systems can often require the development of separate mouse reagents, if the therapeutic agent of interest does not cross-react with mouse. Find out how the AlphaLISA® human PD-1/PD-L1 and AlphaLISA mouse PD-1/PD-L1 binding assays provide a fast, powerful, homogeneous platform for obtain binding potencies from potential novel drug candidates.
Many tumor cells, which under normal, healthy conditions would be recognized by the body’s T cells and thereby targeted for destruction, have developed ways to evade the host immune system by taking advantage of immune checkpoint pathways. Among the most promising approaches to activating therapeutic antitumor immunity is through the blockade of immune checkpoints. The programmed cell death-1 (PD-1) immune checkpoint pathway is a negative regulator of T-cell immune function. When PD-1 is bound to programmed cell death-ligand 1 (PD-L1), T cell response is suppressed. Inhibitors that block PD-1/PD-L1 complex formation lead to increased activation of T-cells and immune system functions, allowing the body’s immune system to identify and attack tumor cells. So far, several anti-PD-1 or PD-L1 monoclonal antibodies have been developed to treat a variety of cancers, including non-small cell lung carcinoma (NSCLC), metastatic melanoma and renal cancer. The promise of therapeutically exploiting this pathway has created a need for more robust, straight-forward assays to identify and qualify potential inhibitors which interrupt PD-1/PD-L1 binding. Find out how AlphaLISA® technology provides a simple, homogenous, straightforward method for detecting PD-1/PD-L1 binding.