Inhibition of SARS‐CoV‐2 spike and ACE2 interaction by thiol molecules was tested using the SARS‐CoV‐2 Spike‐ACE2 interaction Inhibitor Screening Assay kit from Cayman Chemical (MI, USA). To evaluate whether the LMW thiols could disrupt the native form of S1 RBD, the protocol, was slightly modified: the recombinant SARS‐CoV‐2 spike S1 RBD bound to the pre‐coated plate, was incubated with LMW thiols at 37°C for 1 h, and after several washes, the His‐tagged ACE2 protein was added. All the remaining procedures were conducted as indicated by the manufacturer.
Sars cov 2 spike ace2 interaction inhibitor screening assay kit
Manufactured by Cayman Chemical
Sourced in United States
The SARS-CoV-2 Spike-ACE2 Interaction Inhibitor Screening Assay Kit is a tool for detecting compounds that inhibit the interaction between the SARS-CoV-2 spike protein and the human ACE2 receptor. The kit provides a method to screen potential inhibitors of this interaction, which is a key step in the virus entry process.
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4 protocols using sars cov 2 spike ace2 interaction inhibitor screening assay kit
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SARS-CoV-2 Spike-ACE2 Inhibition Assay
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SARS-CoV-2 Spike-ACE2 Inhibition Screening
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A specific SARS-CoV-2 Spike-ACE2 Interaction Inhibitor Screening Assay Kit (Item No. 502050; Cayman Chemical®, Ann Arbor, MI, USA) was used to assess the inhibitory potential of the recombinant SARS-CoV-2 Spro S1 RBD interaction with recombinant ACE-2. Amentoflavone, kaempferitrin, quercetin, luteolin, quercetin-7-O-glucoside (Quercimeritrin) and myricetin were obtained commercially (Sigma-Aldrich St. Louis, MO, USA; analytical grade 93–99%) and evaluated each at concentrations of 0.25; 0.5; 1; 2; 4, and 8 µg/mL. The test was prepared according to the manufacturer’s guidelines and read at 450 nm (Synergy™ H1, Biotek/AGILENT TECHNOLOGIES, Santa Clara, CA, USA).
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SARS-CoV-2 Spike-ACE2 Inhibition Assay
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For evaluating the anti-SARS-CoV-2 activity, the protocol of the SARS-CoV-2 Spike-ACE2 Interaction Inhibitor Screening Assay Kit (Cayman Chemical, USA) was used. A recombinant rabbit Fc-tagged SARS-CoV-2 spike S1 RBD that binds to a plate precoated with a mouse anti-rabbit antibody is used and a recombinant His-tagged ACE2 protein binds to the spike RBD. This complex is detected with an HRP-conjugated anti-His antibody and the absorbance is read at 450 nm. A control is used for competition of the SARS-CoV-2 spike RBD-ACE2 interaction. STD and STE concentrations (0–50 µg-mL) were decided according to the cytotoxicity assay. The activity percentage was calculated according to the given formula:
% Activity = (corrected inhibitor activity) / (corrected 100% initial activity) × 100 (Equation 05).
% Activity = (corrected inhibitor activity) / (corrected 100% initial activity) × 100 (Equation 05).
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SARS-CoV-2 Spike-ACE2 Interaction Assay
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SARS-CoV-2 Spike-ACE2 Interaction Inhibitor Screening Assay Kit (Item No. 502050, Cayman Chemical, USA) was used to evaluate the possible activation effect of AMSIN and emorisin on the RBD binding to ACE2 according to the manufacturer's instructions.
Although this kit was designed to identify inhibitors of the SARS-CoV-2 Spike and ACE2 interaction, quantification by reading the absorbance originating from horseradish peroxidase (HRP) catalysis makes it also suitable to identify an activator. This assay reflects the amount of the enzyme-linked ACE2 captured by a recombinant rabbit Fc-tagged SARS-CoV-2 Spike S1 RBD that binds to a plate precoated with a mouse anti-rabbit antibody. If the amount is increased, one can infer that the peptide activates the ACE2-binding activity of WT RBD.
Although this kit was designed to identify inhibitors of the SARS-CoV-2 Spike and ACE2 interaction, quantification by reading the absorbance originating from horseradish peroxidase (HRP) catalysis makes it also suitable to identify an activator. This assay reflects the amount of the enzyme-linked ACE2 captured by a recombinant rabbit Fc-tagged SARS-CoV-2 Spike S1 RBD that binds to a plate precoated with a mouse anti-rabbit antibody. If the amount is increased, one can infer that the peptide activates the ACE2-binding activity of WT RBD.
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