Spike S1-Biotin (SARS-CoV-2): ACE2 TR-FRET Assay Kit
The pandemic coronavirus disease 2019 (COVID-19) is caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). As a first step of the viral replication strategy, the virus attaches to the host cell surface before entering the cell. The Spike protein recognizes and attaches to the Angiotensin-Converting Enzyme 2 (ACE2) receptor found on the surface of type I and II pneumocytes, endothelial cells, and ciliated bronchial epithelial cells. Drugs targeting the interaction between the Spike protein of SARS-CoV-2 and ACE2 may offer some protection against the viral infection.
The SARS-CoV-2 Spike S1:ACE2 TR-FRET Assay is designed to measure the inhibition of the binding between SARS-CoV-2 Spike S1 and human ACE2 in a homogeneous 96 or 384 reaction format. This TR-FRET-based assay requires no time-consuming washing steps, making it especially suitable for high throughput screening applications. The assay procedure is straightforward and simple; the test inhibitor compound is incubated with biotinylated Spike S1, Eu-labeled ACE2, dye-labeled acceptor and an inhibitor for one hour. Then the TR-FRET signal is measured using a fluorescence reader.
Fluorescence microplate reader capable of measuring Time Resolved-Fluorescence Resonance Energy Transfer (TR-FRET)
Adjustable micropipettor and sterile tips
96 Reactions:
| Catalog Number |
Component |
Amount |
Storage |
|
| 100705 | ACE2, His-Tag, Eu-labeled* | 2 µg | -80°C | Avoid multiple freeze/ thaw cycles! |
| 100679 | Spike S1, Fc fusion, Avi-tag, Biotin-Labeled (SARS-CoV-2)* | 25 µg | -80°C | |
| Dye-labeled acceptor | 2x10 µl | -20°C | ||
| 79953 | 3x ACE2-Spike TR-FRET Buffer | 4 ml | -20°C | |
| 79696 | 96-well white microplate | 1 | Room Temp |
|
*The concentration of ACE2 and Spike S1 is lot-specific and will be indicated on the tubes containing the enzyme.
384 Reactions:
| Catalog Number |
Component |
Amount |
Storage |
|
| 100705 | ACE2, His-Tag, Eu-labeled* | 2x2 µg | -80°C | Avoid multiple freeze/ thaw cycles! |
| 100679 | Spike S1, Fc fusion, Avi-tag, Biotin-Labeled (SARS-CoV-2)* | 2x25 µg | -80°C | |
| Dye-labeled acceptor | 3x10 µl | -20°C | ||
| 79953 | 3x ACE2-Spike TR-FRET Buffer | 4 ml | -20°C | |
| 384-well white microplate | 1 | Room Temp |
||
*The concentration of ACE2 and Spike S1 is lot-specific and will be indicated on the tubes containing the enzyme.
Hoffmann, M. et al. 2020. Cell, 181:1-10
Yan, R. et al. Science, 4 March 2020. In Press. DOI: 10.1126/science.abb2762
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