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Spike S1 (B.1.1.7, Alpha Variant) (SARS-CoV-2) : ACE2 Inhibitor Screening Colorimetric Assay Kit

The Spike S1 (B.1.1.7 Variant) (SARS-CoV-2) :ACE2 Inhibitor Screening Colorimetric Assay Kit is designed for screening and profiling inhibitors of the interaction of ACE2 (angiotensin-converting enzyme 2) with the B.1.1.7 variant of the SARS-CoV-2 Spike S1 protein. This kit comes in a convenient 96-well format, with purified SARS-CoV-2 Spike S1 variant B1.1.7 and ACE2-Biotin (amino acids 18-740) proteins, streptavidin-HRP, colorimetric HRP substrate, and assay buffer for 100 binding reactions. The SARS-CoV-2 Spike S1 (B.1.1.7 Variant) protein, included in the kit provides a biologically relevant model for the investigation of SARS-CoV-2/ACE2 interaction.Figure 1. Assay Kit schematic.First, SARS-CoV-2 Spike S1 (B.1.1.7 Variant) is coated on a 96-well plate overnight. After washing and blocking, the protein is pre-incubated with an inhibitor or neutralizing antibody. Upon subsequent incubation with Biotin-ACE2, the plate is treated with Streptavidin-HRP. After a final wash, Colorimetric HRP substrate is added to produce color that can be measured using a UV/Vis spectrophotometer microplate reader. The colorimetric signal is proportional to the efficacy of the binding of Spike S1 (B.1.1.7 Variant) to ACE2.Interested in having us perform the screening and profiling of inhibitors or blocking antibodies? Visit ourCoronavirus Service page.

Product Specifications

Background

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 of the SARS-CoV-2 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. It has been widely suggested that active as well as passive immunizations targeting the interaction between the Spike protein of SARS-CoV-2 and ACE2 offer promising protection against the viral infection. Reports showed that a mutant strain first identified in the UK (B1.1.7) exhibits higher transmissibility and infectivity. The B.1.1.7 variant contains multiple mutations, including several in the Spike protein that leads to higher infectivity rates than the wild-type virus. The S1 subunit (a.a. 14-685) of the Spike protein includes the Receptor Binding Domain (RBD) region (a.a. 319-591) that is responsible for binding to the ACE2 receptor on target cells.  Mutations outside of the RBD in the S1 subunit of spike are important for influencing the immunogenicity, conformation, and flexibility of the spike protein.  Investigations on the effects of mutations on viral replication and pathogenesis will be critical for developing effective strategies for vaccines and antibody therapies against COVID-19.

Applications

Screening inhibitors of ACE2 binding to SARS-CoV-2 Spike S1 B.1.1.7 Variant.

Format

Catalog #NameAmountStorageSpike S1 (del_HV69-70, del_Y144, N501Y, A570D, D614G, P681H), His-tag (SARS-CoV-2) 10 µg-80°C100665ACE2, His-Avi-Tag, Biotin-labeled HiP™5 µg-80°C79742Streptavidin-HRP10 µl+4°C793113x Immuno Buffer 150 ml-20°C79728Blocking Buffer 250 ml+4°C79651Colorimetric HRP substrate10 ml+4°C79964Transparent 96-well microplate1Room Temp

Shipping Conditions

-80°C

Storage Conditions

This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.

Notes

Visitbpsbioscience.com/assay-kits-faqfor detailed troubleshooting instructions. For all further questions, please email[email protected]

Contraindications

This assay kit is compatible with up to 1% final DMSO concentration.

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