Spike S1 RBD (B.1.351, Beta Variant) (SARS-CoV-2) : ACE2 [Biotinylated] Inhibitor Screening Colorimetric Assay Kit

• 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 Receptor Binding Domain (RBD) of 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. 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. A variant strain of SARS-COV-2, identified as B.1.351, was first identified in the fall of 2020 in the Republic of South Africa. This South African variant, also known as 501Y.V2, has many mutations which may lead to higher transmissibility and infectivity.  In particular, the B.1.351 variant contains three mutations within the RBD region of the Spike S1 protein: K417N, E484K, and N501Y.  Investigating the effect of mutations on viral replication and pathogenesis will be critical for developing effective strategies for vaccines and antibody therapies against COVID-19.

• Description:

  The Spike S1 RBD-B.1.351 (SARS-CoV-2) :ACE2 [Biotinylated] Inhibitor Screening Colorimetric Assay Kit is designed for screening and profiling inhibitors of the interaction of ACE2 (angiotensin-converting enzyme 2) with the RBD (receptor binding domain) region of the B.1.351 Variant of the SARS-CoV-2 Spike S1 protein. This kit comes in a convenient 96-well format, with purified Spike S1 RBD-B.1.351 (SARS-CoV-2) (amino acids 319-541) and ACE2-Biotin (amino acids 18-740) proteins, streptavidin-HRP, colorimetric HRP substrate, and assay buffer for 100 binding reactions. The Spike S1 RBD-B.1.351 (SARS-CoV-2) 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, Spike S1 RBD-B.1.351 (SARS-CoV-2) 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 RBD-B.1.351 (SARS-CoV-2) to ACE2.Interested in having us perform the screening and profiling of inhibitors or blocking antibodies? Visit ourCoronavirus Screening Services.

• Applications:

  Screening inhibitors of ACE2 binding to Spike S1 RBD-B.1.351 (SARS-CoV-2) .

• Format:

  Catalog #NameAmountStorage100978Spike RBD (B.1.351, Beta Variant) Avi-His-Tag (SARS-CoV-2) *5 µg-80°C100665ACE2, His-Avi-Tag, Biotin-Labeled*5 µg-80°C793113x Immuno Buffer 150 ml-20°C79728Blocking Buffer 250 ml+4°C79742Streptavidin-HRP10 µl+4°C79651Colorimetric HRP Substrate10 ml+4°C79964Transparent 96-well microplate1Room Temp*The initial concentration of the proteins is lot-specific and will be indicated on the tube containing the protein.

• 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.