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TNFα/TNFR1 (human) AlphaLISA Binding Kit, 500 Assay Points

The AlphaLISA® human TNFα/TNFR1 binding kit is designed for the detection of binding activity between human TNFα and TNFR1, 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.

For research use only. Not for use in diagnostic procedures.

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AL3120C
500 Assay Points
2213.00 USD
 
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AL3120F
5,000 Assay Points
11400.00 USD
 
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Overview

The AlphaLISA® human TNFα/TNFR1 binding assay uses anti-6xHis AlphaLISA Acceptor beads to capture the His-tagged TNFR1 and Streptavidin-coated donor beads to capture the biotinylated TNFα. Donor beads and Acceptor beads come into proximity through TNFα binding to TNFR1. 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.

Formats:

  • Our 500 assay point kit allows you to run 500 wells in 96-well or 384-well format, using a 20 µL reaction volume.
  • Our 5,000 assay point kit allows you to run 5,000 wells in 96-well or 384-well format, using a 20 µL reaction volume.

AlphaLISA features:

  • No-wash steps, no separation steps
  • Ease-of-use: few addition steps, fast assay development
  • Broad range of affinities: detect strong or weak interactions, from pM to mM affinity
  • Distance: measure very large protein or antibody complexes – spanning up to 200 nm or more
  • High avidity: multiple binding sites on each bead enables use of nanomolar concentrations of antibodies or proteins, as well as use of low affinity binders

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)]. TNFR1 is ubiquitously expressed and can be fully activated by both the membrane-bound and soluble trimeric forms of TNFα, whereas TNFR2 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.

Specifications

Assay Target TNFα,TNRFR1
Assay Target Class Cytokine, Protein
Automation Compatible Yes
Detection Method Alpha
Product Brand Name AlphaLISA
Shipping Condition Blue Ice
Therapeutic Area Inflammation
Unit Size 500 Assay Points
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Application Note

Evaluating the Specificity of PD-1 and PD-L1 Blocking Antibodies Using AlphaLISA Human and Mouse PD-1/PD-L1 Binding Kits

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.

PDF 1 MB
Screening for Inhibitors of PD-1 and PD-L1 Binding with AlphaLISA Technology

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.

PDF 1 MB

Brochure

Alpha Technology Solutions

Product brochure for the Alpha Technology, a versatile, no wash, homogeneous assay technology that's suitable for a broad range of applications.

PDF 4 MB