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Angiotensin-converting enzyme 2 (ACE2) is a human cell surface receptor that has been identified as the main point of entry for SARS-CoV-2 virions, which match and bind ACE2 with their own viral Spike protein S1 subunits. This interaction is therefore a key target in COVID-19 therapeutic research, as its inhibitors may yield a protective effect by preventing SARS-CoV-2 entry into cells. For this reason, protein-protein interaction assays suitable for ACE2-Spike inhibitor screening are relevant to current research.
The AlphaLISA detection of SARS-CoV-2 S1/ACE2 binding uses acceptor beads coupled to human ACE2 protein and Streptavidin-coated donor beads to capture the biotinylated S1-protein. Donor beads and acceptor beads come into proximity through S1 binding to ACE2. 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 emission at 615 nm (Figure 1).This assay can facilitate the screening of inhibitors of S1/ACE2 binding. A competitor of the interaction will disrupt the S1/ACE2 binding leading to a decrease of the AlphaLISA Signal.
|Assay Target||SARS-CoV2 ACE2/S1|
|Experimental Type||In vitro|
|Product Brand Name||AlphaLISA|
|Shipping Condition||Blue Ice|
|Unit Size||5,000 Assay Points|
Alpha (Amplified Luminescent Proximity Homogeneous Assay) technology is a bead-based, no-wash alternative to traditional ELISAs. Instead of detection with an HRP substrate, the Alpha assay signal is generated by the excitation of an Eu+-coated bead that has been conjugated to the detection antibody.
Alpha technology offers a simple, straight forward workflow. No wash needed!
Download this application note to see how no-wash AlphaLISA® technology provides a more-convenient alternative to ELISA for quantitation of biomarkers in complex sample types, including tissue, serum, and plasma.
Biomarker levels were measured directly in CulturPlates-96 and-384 in a simple, fast, all-in-one-well AlphaLISA assay format. The elimination of transfer and wash steps simplifies cellular assays, reduces variability and significantly reduces hands-on time and costs associated with consumables. Integral plasma membrane (EGFR) and secreted solubleproteins (TNFa, IL1ß, IL6, IL8) were successfully determined, on adherent or suspension cells, using the standard AlphaLISA Immunoassay buffer.
The AlphaLISA® assay is a homogeneous immunoassay alternative to classical ELISA. AlphaLISA assays were originally utilized to detect analytesin cell cultures upernatants or serum/plasma samples.