SARS-CoV-1 Spike Trimer (S1+S2):ACE2 Inhibitor Screening Colorimetric Assay Kit
Severe acute respiratory syndrome (SARS) is a viral respiratory illness caused by a corona virus called SARS-CoV-1. It was first reported in February 2013 in Asia and the epidemic affected ~ 26 countries and resulted in more than 8000 deaths. Like SARS-CoV-2, SARS-CoV-1 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-1 and ACE2 may offer some protection against the viral infection. The SARS-CoV-1 Spike Trimer (S1+S2):ACE2 Inhibitor Screening Assay Kit includes the SARS-CoV-1 Spike protein in its native trimeric conformation to provide a more physiologically relevant screen for inhibitors as well as a compatible platform to investigate the specificity of SARS-CoV-2:ACE2 inhibitors.
The key to this kit is that the SARS-CoV-1 Spike Trimer (S1+S2) protein provides a more biologically relevant model than monomeric Spike RBD protein for the investigation of SARS-CoV-1/host cell interaction. Only a few simple steps on a microtiter plate are required for the assay. First, SARS-CoV-1 Spike Trimer (S1+S2) is coated on a 96-well plate. Next, Biotin-ACE2 is incubated with SARS-CoV-1 Spike Trimer (S1+S2) on the plate. Finally, the plate is treated with Streptavidin-HRP followed by addition of a colorimetric HRP substrate to produce color, which then can be quenched and measured using a UV/Vis microplate reader.
Assay Principle
This kit is for detection of inhibitors of the Spike trimers from SARS-COV-1, the virus responsible for Severe Acute Respiratory Syndrome (SARS) disease, not SARS-COV-2, the causative agent of COVID-19. To detect inhibitors of the SARS-COV-2 Spike trimer, please use SARS-CoV-2 Spike Trimer (S1+S2):ACE2 Inhibitor Screening Colorimetric Assay Kit (BPS Bioscience #79999).
PBS (Phosphate buffered saline)
PBST (Phosphate buffered saline containing 0.05% Tween-20)
1N HCl (aqueous)
Rotating or rocker platform
UV/Vis spectrophotometer microplate reader capable of reading absorbance at 450 nm*
| Catalog Number |
Component |
Amount |
Storage |
|
| 100789 | Spike Trimer (S1 + S2), His-Tag (SARS-CoV-1)* | 5 µg | -80°C | Avoid multiple freeze/ thaw cycles! |
| 100665 | ACE2, His-Avi-Tag, Biotin labeled* | 5 µg | -80°C | |
| 79311 | 3x Immuno Buffer 1 | 50 ml | -20°C | |
| 79728 | Blocking Buffer 2 | 50 ml | +4°C | |
| 79742 | Streptavidin-HRP | 5 µl | +4°C | |
| Colorimetric HRP substrate | 10 ml | +4°C | ||
| 79964 | Transparent 96-well white microplate | 1 | Room Temp |
|
*The initial concentration of spike trimer and ACE2 is lot-specific and will be indicated on the tube containing the proteins.
Yan, R. et al. 2020. Science 367 (6485): 1444-1448.
Li, W., et al. 2003. Angiotensin-converting enzyme 2 is a functional receptor for the SARS coronavirus. Nature 426:450-454.
Li, J., et al. 2013. Immunogenicity and Protection Efficacy of Monomeric and Trimeric Recombinant SARS Coronavirus Spike Protein Subunit Vaccine Candidates. Viral Immunol. 26(2): 126–132.
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