sars cov 2 spike protein mice fc chimera  (Sino Biological)


Bioz Verified Symbol Sino Biological is a verified supplier
Bioz Manufacturer Symbol Sino Biological manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 96
    Name:
    SARS CoV 2 2019 nCoV Spike RBD Fc Recombinant Protein HPLC verified COVID 19 Spike RBD Research
    Description:
    A DNA sequence encoding the SARS CoV 2 2019 nCoV Spike Protein RBD YP 009724390 1 Arg319 Phe541 was expressed with the Fc region of human IgG1 at the C terminus
    Catalog Number:
    40592-V02H
    Price:
    None
    Category:
    recombinant protein
    Product Aliases:
    coronavirus spike Protein 2019-nCoV, cov spike Protein 2019-nCoV, ncov RBD Protein 2019-nCoV, ncov s1 Protein 2019-nCoV, ncov s2 Protein 2019-nCoV, ncov spike Protein 2019-nCoV, NCP-CoV RBD Protein 2019-nCoV, NCP-CoV s1 Protein 2019-nCoV, NCP-CoV s2 Protein 2019-nCoV, NCP-CoV Spike Protein 2019-nCoV, novel coronavirus RBD Protein 2019-nCoV, novel coronavirus s1 Protein 2019-nCoV, novel coronavirus s2 Protein 2019-nCoV, novel coronavirus spike Protein 2019-nCoV, RBD Protein 2019-nCoV, S1 Protein 2019-nCoV, S2 Protein 2019-nCoV, Spike RBD Protein 2019-nCoV
    Host:
    HEK293 Cells
    Buy from Supplier


    Structured Review

    Sino Biological sars cov 2 spike protein mice fc chimera
    Binding of <t>SARS-CoV-2</t> spike protein to different S. pneumoniae CPS. Wells were coated with different S. pneumoniae CPS and the binding to the viral spike protein was evaluated exploiting the mouse-Fc tag on the recombinant chimera spike protein. The experiment has been performed three times in duplicate with similar results, and data were normalized over signal from blocking buffer-coated wells. Error bars indicate standard deviations. OD: Optical density.
    A DNA sequence encoding the SARS CoV 2 2019 nCoV Spike Protein RBD YP 009724390 1 Arg319 Phe541 was expressed with the Fc region of human IgG1 at the C terminus
    https://www.bioz.com/result/sars cov 2 spike protein mice fc chimera/product/Sino Biological
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 spike protein mice fc chimera - by Bioz Stars, 2021-04
    96/100 stars

    Images

    1) Product Images from "Novel ACE2-Independent Carbohydrate-Binding of SARS-CoV-2 Spike Protein to Host Lectins and Lung Microbiota"

    Article Title: Novel ACE2-Independent Carbohydrate-Binding of SARS-CoV-2 Spike Protein to Host Lectins and Lung Microbiota

    Journal: bioRxiv

    doi: 10.1101/2020.05.13.092478

    Binding of SARS-CoV-2 spike protein to different S. pneumoniae CPS. Wells were coated with different S. pneumoniae CPS and the binding to the viral spike protein was evaluated exploiting the mouse-Fc tag on the recombinant chimera spike protein. The experiment has been performed three times in duplicate with similar results, and data were normalized over signal from blocking buffer-coated wells. Error bars indicate standard deviations. OD: Optical density.
    Figure Legend Snippet: Binding of SARS-CoV-2 spike protein to different S. pneumoniae CPS. Wells were coated with different S. pneumoniae CPS and the binding to the viral spike protein was evaluated exploiting the mouse-Fc tag on the recombinant chimera spike protein. The experiment has been performed three times in duplicate with similar results, and data were normalized over signal from blocking buffer-coated wells. Error bars indicate standard deviations. OD: Optical density.

    Techniques Used: Binding Assay, Recombinant, Blocking Assay

    Binding of SARS-CoV-2 spike protein to different LPS. Wells were coated with different LPS and the binding to the viral spike protein was evaluated exploiting the mouse-Fc tag on the recombinant chimera spike protein. The experiment has been performed three times in duplicate with similar results, and data were normalized over signal from blocking buffer-coated wells. Error bars indicate standard deviations. OD: Optical density.
    Figure Legend Snippet: Binding of SARS-CoV-2 spike protein to different LPS. Wells were coated with different LPS and the binding to the viral spike protein was evaluated exploiting the mouse-Fc tag on the recombinant chimera spike protein. The experiment has been performed three times in duplicate with similar results, and data were normalized over signal from blocking buffer-coated wells. Error bars indicate standard deviations. OD: Optical density.

    Techniques Used: Binding Assay, Recombinant, Blocking Assay

    Binding of SARS-CoV-2 spike protein to human C-type lectins. Human C-type lectins ELISA results: wells were coated with the viral spike protein and the binding to different C-type lectins was evaluated in a calcium-containing buffer. The experiment has been performed three times in duplicate with similar results, and data were normalized over signal from BSA-coated wells. Error bars indicate standard deviations. OD: Optical density.
    Figure Legend Snippet: Binding of SARS-CoV-2 spike protein to human C-type lectins. Human C-type lectins ELISA results: wells were coated with the viral spike protein and the binding to different C-type lectins was evaluated in a calcium-containing buffer. The experiment has been performed three times in duplicate with similar results, and data were normalized over signal from BSA-coated wells. Error bars indicate standard deviations. OD: Optical density.

    Techniques Used: Binding Assay, Enzyme-linked Immunosorbent Assay

    Binding of SARS-CoV-2 spike protein to human Siglecs. Human Siglecs ELISA results: wells were coated with the viral spike protein and the binding to different Siglces was evaluated. The experiment has been performed three times in duplicate with similar results, and data were normalized over signal from blocking buffer-coated wells. Error bars indicate standard deviations. OD: Optical density.
    Figure Legend Snippet: Binding of SARS-CoV-2 spike protein to human Siglecs. Human Siglecs ELISA results: wells were coated with the viral spike protein and the binding to different Siglces was evaluated. The experiment has been performed three times in duplicate with similar results, and data were normalized over signal from blocking buffer-coated wells. Error bars indicate standard deviations. OD: Optical density.

    Techniques Used: Binding Assay, Enzyme-linked Immunosorbent Assay, Blocking Assay

    2) Product Images from "Targeting ACE2–RBD Interaction as a Platform for COVID-19 Therapeutics: Development and Drug-Repurposing Screen of an AlphaLISA Proximity Assay"

    Article Title: Targeting ACE2–RBD Interaction as a Platform for COVID-19 Therapeutics: Development and Drug-Repurposing Screen of an AlphaLISA Proximity Assay

    Journal: ACS Pharmacology & Translational Science

    doi: 10.1021/acsptsci.0c00161

    Scheme describing the assays employed in this paper. (A) The recombinant protein constructs ACE2-His-Avi (ACE2-Avi) (Acro Biosystems) and SARS-CoV-2 spike protein receptor binding domain-Fc (RBD-Fc) (Sino Biological) were used to model ACE2–RBD binding. (B) AlphaLISA assay system used to monitor ACE2–RBD interacts. Streptavidin donor beads recognize the Avi tag on ACE2. The protein A acceptor beads recognize the Fc tag on RBD. When in proximity the donor beads can be excited with light at 680 nm. This generates singlet oxygen which diffuses to the acceptor beads, causing the acceptor beads to luminesce at 615 nm. (C) The TruHits counterscreen uses streptavidindonor beads which directly interact with biotin acceptor beads. Because no intermediary molecule is needed to bring the donor and acceptor beads in proximity, the TruHits assay can identify compounds which directly interfere with the AlphaLISA readout.
    Figure Legend Snippet: Scheme describing the assays employed in this paper. (A) The recombinant protein constructs ACE2-His-Avi (ACE2-Avi) (Acro Biosystems) and SARS-CoV-2 spike protein receptor binding domain-Fc (RBD-Fc) (Sino Biological) were used to model ACE2–RBD binding. (B) AlphaLISA assay system used to monitor ACE2–RBD interacts. Streptavidin donor beads recognize the Avi tag on ACE2. The protein A acceptor beads recognize the Fc tag on RBD. When in proximity the donor beads can be excited with light at 680 nm. This generates singlet oxygen which diffuses to the acceptor beads, causing the acceptor beads to luminesce at 615 nm. (C) The TruHits counterscreen uses streptavidindonor beads which directly interact with biotin acceptor beads. Because no intermediary molecule is needed to bring the donor and acceptor beads in proximity, the TruHits assay can identify compounds which directly interfere with the AlphaLISA readout.

    Techniques Used: Recombinant, Construct, Binding Assay

    Related Articles

    Transfection:

    Article Title: Experimental data using candesartan and captopril indicate no double-edged sword effect in COVID-19
    Article Snippet: Cells were transiently transfected with 2 μg of ACE2 cDNA (ACE2 tGFP-tagged, RG208442, Origene) using a commercial transfection reagent, Turbofect (R0533, Thermo Scientific). .. SARS-CoV-2 Spike RBD-Fc protein internalization assayHuman alveolar type-II pneumocyte cell lines, A549, transiently transfected with tGFP ACE2 were treated or not treated with Candesartan or Captopril for 24 h. Then, 1 µg/ml of SARS-CoV-2 Spike RBD-Fc protein (40592-V02H, Sino Biological) was added to the cells for 3 h at 37°C. .. Cells were fixed and incubated overnight at 4°C with a mouse monoclonal antibody against human IgG-Fc (ab99757, abcam, 1:500) diluted in DPBS containing 1% BSA, 2% normal goat serum and 0.05% Triton X-100.

    Recombinant:

    Article Title: Androgen Regulates SARS-CoV-2 Receptor Levels and Is Associated with Severe COVID-19 Symptoms in Men
    Article Snippet: .. After 24 hours, 0.5 μM/mL human Fc-tagged recombinant spike-RBD protein with (Sino Biological Inc. 40592-V02H) was added to the medium and incubated for 30 minutes. .. The cells were subsequently fixed, permeabilized and stained with antibodies against ACE2 and human Fc receptor.

    Article Title: Androgen Signaling Regulates SARS-CoV-2 Receptor Levels and Is Associated with Severe COVID-19 Symptoms in Men
    Article Snippet: .. After 24 h, 0.5 μM/mL human Fc-tagged recombinant spike-RBD protein (Sino Biological Inc. 40592-V02H) was added to the medium and incubated for 30 min. .. The cells were subsequently fixed, permeabilized, and stained with antibodies against ACE2 and human Fc receptor, as described previously, followed by high-throughput imaging and quantification using the In Cell Analyzer 2000 (GE Healthcare, USA).

    Incubation:

    Article Title: Androgen Regulates SARS-CoV-2 Receptor Levels and Is Associated with Severe COVID-19 Symptoms in Men
    Article Snippet: .. After 24 hours, 0.5 μM/mL human Fc-tagged recombinant spike-RBD protein with (Sino Biological Inc. 40592-V02H) was added to the medium and incubated for 30 minutes. .. The cells were subsequently fixed, permeabilized and stained with antibodies against ACE2 and human Fc receptor.

    Article Title: Androgen Signaling Regulates SARS-CoV-2 Receptor Levels and Is Associated with Severe COVID-19 Symptoms in Men
    Article Snippet: .. After 24 h, 0.5 μM/mL human Fc-tagged recombinant spike-RBD protein (Sino Biological Inc. 40592-V02H) was added to the medium and incubated for 30 min. .. The cells were subsequently fixed, permeabilized, and stained with antibodies against ACE2 and human Fc receptor, as described previously, followed by high-throughput imaging and quantification using the In Cell Analyzer 2000 (GE Healthcare, USA).

    Blocking Assay:

    Article Title: Novel ACE2-Independent Carbohydrate-Binding of SARS-CoV-2 Spike Protein to Host Lectins and Lung Microbiota
    Article Snippet: After discarding and washing (2×150µL) with PBS (10mM, pH=7.4), the wells were blocked with 100 µL of carbo-free blocking solution (Vector Laboratories, catalog No.NC9977573) at 37°C for 30 min. .. The blocking solution was discarded and 50µL of SARS-CoV-2 spike protein mice-Fc chimera (2019-nCoV Spike Protein RBD, Fc Tag, purchased from Sinobiological) at 2 µg/mL in carbo-free buffer were added to the wells. .. After 1h at room temperature, the wells were washed with PBS (2×150µL) and then 100 µL of anti-mouse horseradish peroxidase (1 µg/mL, Goat anti-mouse IgG-HRP from JacksonImmuno) were added.

    Mouse Assay:

    Article Title: Novel ACE2-Independent Carbohydrate-Binding of SARS-CoV-2 Spike Protein to Host Lectins and Lung Microbiota
    Article Snippet: After discarding and washing (2×150µL) with PBS (10mM, pH=7.4), the wells were blocked with 100 µL of carbo-free blocking solution (Vector Laboratories, catalog No.NC9977573) at 37°C for 30 min. .. The blocking solution was discarded and 50µL of SARS-CoV-2 spike protein mice-Fc chimera (2019-nCoV Spike Protein RBD, Fc Tag, purchased from Sinobiological) at 2 µg/mL in carbo-free buffer were added to the wells. .. After 1h at room temperature, the wells were washed with PBS (2×150µL) and then 100 µL of anti-mouse horseradish peroxidase (1 µg/mL, Goat anti-mouse IgG-HRP from JacksonImmuno) were added.

    Clone Assay:

    Article Title: Antibodies that potently inhibit or enhance SARS-CoV-2 spike protein-ACE2 interaction isolated from synthetic single-chain antibody libraries
    Article Snippet: The library was panned using either SARS-CoV-2 RBD (Sino Biological, Cat: 40592-V08H; screens RU167 and RU169) or soluble S1 ectodomain trimer (Sino Biological, Cat: 40591-V08H; RU171) that was biotinylated using ChromaLINK Biotin (Vector Laboratories, Cat: B-1001) and attached to MyOne Streptavidin C1 Dynabeads (ThermoFisher, Cat: 65002). .. RBD or S1-binding clones were moved into the ReD cell-display platform (Beasley et al. 2015) and cells were membrane permeabilized using 0.5% n-octyl b-d-thioglucopyranoside (Anatrace, Cat: 0314) and labeled for FACS using either SARS-CoV-2 RBD-Fc (Sino Biological, Cat: 40592-V02H; screens RU167 and RU169) or soluble S1 ectodomain trimer (Sino Biological, Cat: 40591-V08H; RU171) that had been fluorophore labeled with either ATTO 488 NHS ester (Merck, Cat: 41698) or Dy-549P1 (Dyomics, Cat: 549P1-01). ..

    Labeling:

    Article Title: Antibodies that potently inhibit or enhance SARS-CoV-2 spike protein-ACE2 interaction isolated from synthetic single-chain antibody libraries
    Article Snippet: The library was panned using either SARS-CoV-2 RBD (Sino Biological, Cat: 40592-V08H; screens RU167 and RU169) or soluble S1 ectodomain trimer (Sino Biological, Cat: 40591-V08H; RU171) that was biotinylated using ChromaLINK Biotin (Vector Laboratories, Cat: B-1001) and attached to MyOne Streptavidin C1 Dynabeads (ThermoFisher, Cat: 65002). .. RBD or S1-binding clones were moved into the ReD cell-display platform (Beasley et al. 2015) and cells were membrane permeabilized using 0.5% n-octyl b-d-thioglucopyranoside (Anatrace, Cat: 0314) and labeled for FACS using either SARS-CoV-2 RBD-Fc (Sino Biological, Cat: 40592-V02H; screens RU167 and RU169) or soluble S1 ectodomain trimer (Sino Biological, Cat: 40591-V08H; RU171) that had been fluorophore labeled with either ATTO 488 NHS ester (Merck, Cat: 41698) or Dy-549P1 (Dyomics, Cat: 549P1-01). ..

    FACS:

    Article Title: Antibodies that potently inhibit or enhance SARS-CoV-2 spike protein-ACE2 interaction isolated from synthetic single-chain antibody libraries
    Article Snippet: The library was panned using either SARS-CoV-2 RBD (Sino Biological, Cat: 40592-V08H; screens RU167 and RU169) or soluble S1 ectodomain trimer (Sino Biological, Cat: 40591-V08H; RU171) that was biotinylated using ChromaLINK Biotin (Vector Laboratories, Cat: B-1001) and attached to MyOne Streptavidin C1 Dynabeads (ThermoFisher, Cat: 65002). .. RBD or S1-binding clones were moved into the ReD cell-display platform (Beasley et al. 2015) and cells were membrane permeabilized using 0.5% n-octyl b-d-thioglucopyranoside (Anatrace, Cat: 0314) and labeled for FACS using either SARS-CoV-2 RBD-Fc (Sino Biological, Cat: 40592-V02H; screens RU167 and RU169) or soluble S1 ectodomain trimer (Sino Biological, Cat: 40591-V08H; RU171) that had been fluorophore labeled with either ATTO 488 NHS ester (Merck, Cat: 41698) or Dy-549P1 (Dyomics, Cat: 549P1-01). ..

    other:

    Article Title: Targeting ACE2–RBD Interaction as a Platform for COVID-19 Therapeutics: Development and Drug-Repurposing Screen of an AlphaLISA Proximity Assay
    Article Snippet: RBD-Fc (SARS-CoV-2 spike protein residues 319–541, C-terminal Fc tag; catalog no.40592-V02H), ACE2-His (ACE2 residues 1–740, C-terminal His tag; catalog no. 10108-H08H), and S1-His (spike protein residues 1–667, C-terminal His tag; catalog no. 40150-V08B1) were acquired from Sino Biological (Wayne, PA).

    Article Title: Targeting ACE2-RBD interaction as a platform for COVID19 therapeutics: Development and drug repurposing screen of an AlphaLISA proximity assay
    Article Snippet: RBD-Fc (SARS-CoV-2 spike protein residues 319-541, C-terminal Fc tag) (Cat #40592-V02H), ACE2-His (ACE2 residues 1-740, C-terminal His tag) (Cat # 10108-H08H), and S1-His (spike protein residues 1-667, C-terminal His tag) (Cat # 40150-V08B1) were acquired from Sino Biological (Wayne, PA).

    Binding Assay:

    Article Title: Detection of SARS-CoV-2 neutralizing antibodies with a cell-free PCR assay
    Article Snippet: The SARS-CoV-2 spike protein (S1) containing amino acids 1-674 with an Fc-tag at the C-terminus (#31806) expressed in HEK293 cells was purchased from the Native Antigen Company (Oxford, United Kingdom). .. The SARS-CoV-2 spike protein receptor binding domain (RBD) containing amino acids 319-541 with an Fc-tag at the Cterminus (#40592-V02H) and the human receptor angiotensin-converting enzyme 2 (ACE2) protein containing amino acids 1-740 with an Fc-tag at the C-terminus (10108- H05H) expressed in HEK293 cells were obtained from Sino Biologicals (Beijing, China). ..

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 96
    Sino Biological sars cov 2 spike protein mice fc chimera
    Binding of <t>SARS-CoV-2</t> spike protein to different S. pneumoniae CPS. Wells were coated with different S. pneumoniae CPS and the binding to the viral spike protein was evaluated exploiting the mouse-Fc tag on the recombinant chimera spike protein. The experiment has been performed three times in duplicate with similar results, and data were normalized over signal from blocking buffer-coated wells. Error bars indicate standard deviations. OD: Optical density.
    Sars Cov 2 Spike Protein Mice Fc Chimera, supplied by Sino Biological, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sars cov 2 spike protein mice fc chimera/product/Sino Biological
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 spike protein mice fc chimera - by Bioz Stars, 2021-04
    96/100 stars
      Buy from Supplier

    Image Search Results


    Binding of SARS-CoV-2 spike protein to different S. pneumoniae CPS. Wells were coated with different S. pneumoniae CPS and the binding to the viral spike protein was evaluated exploiting the mouse-Fc tag on the recombinant chimera spike protein. The experiment has been performed three times in duplicate with similar results, and data were normalized over signal from blocking buffer-coated wells. Error bars indicate standard deviations. OD: Optical density.

    Journal: bioRxiv

    Article Title: Novel ACE2-Independent Carbohydrate-Binding of SARS-CoV-2 Spike Protein to Host Lectins and Lung Microbiota

    doi: 10.1101/2020.05.13.092478

    Figure Lengend Snippet: Binding of SARS-CoV-2 spike protein to different S. pneumoniae CPS. Wells were coated with different S. pneumoniae CPS and the binding to the viral spike protein was evaluated exploiting the mouse-Fc tag on the recombinant chimera spike protein. The experiment has been performed three times in duplicate with similar results, and data were normalized over signal from blocking buffer-coated wells. Error bars indicate standard deviations. OD: Optical density.

    Article Snippet: The blocking solution was discarded and 50µL of SARS-CoV-2 spike protein mice-Fc chimera (2019-nCoV Spike Protein RBD, Fc Tag, purchased from Sinobiological) at 2 µg/mL in carbo-free buffer were added to the wells.

    Techniques: Binding Assay, Recombinant, Blocking Assay

    Binding of SARS-CoV-2 spike protein to different LPS. Wells were coated with different LPS and the binding to the viral spike protein was evaluated exploiting the mouse-Fc tag on the recombinant chimera spike protein. The experiment has been performed three times in duplicate with similar results, and data were normalized over signal from blocking buffer-coated wells. Error bars indicate standard deviations. OD: Optical density.

    Journal: bioRxiv

    Article Title: Novel ACE2-Independent Carbohydrate-Binding of SARS-CoV-2 Spike Protein to Host Lectins and Lung Microbiota

    doi: 10.1101/2020.05.13.092478

    Figure Lengend Snippet: Binding of SARS-CoV-2 spike protein to different LPS. Wells were coated with different LPS and the binding to the viral spike protein was evaluated exploiting the mouse-Fc tag on the recombinant chimera spike protein. The experiment has been performed three times in duplicate with similar results, and data were normalized over signal from blocking buffer-coated wells. Error bars indicate standard deviations. OD: Optical density.

    Article Snippet: The blocking solution was discarded and 50µL of SARS-CoV-2 spike protein mice-Fc chimera (2019-nCoV Spike Protein RBD, Fc Tag, purchased from Sinobiological) at 2 µg/mL in carbo-free buffer were added to the wells.

    Techniques: Binding Assay, Recombinant, Blocking Assay

    Binding of SARS-CoV-2 spike protein to human C-type lectins. Human C-type lectins ELISA results: wells were coated with the viral spike protein and the binding to different C-type lectins was evaluated in a calcium-containing buffer. The experiment has been performed three times in duplicate with similar results, and data were normalized over signal from BSA-coated wells. Error bars indicate standard deviations. OD: Optical density.

    Journal: bioRxiv

    Article Title: Novel ACE2-Independent Carbohydrate-Binding of SARS-CoV-2 Spike Protein to Host Lectins and Lung Microbiota

    doi: 10.1101/2020.05.13.092478

    Figure Lengend Snippet: Binding of SARS-CoV-2 spike protein to human C-type lectins. Human C-type lectins ELISA results: wells were coated with the viral spike protein and the binding to different C-type lectins was evaluated in a calcium-containing buffer. The experiment has been performed three times in duplicate with similar results, and data were normalized over signal from BSA-coated wells. Error bars indicate standard deviations. OD: Optical density.

    Article Snippet: The blocking solution was discarded and 50µL of SARS-CoV-2 spike protein mice-Fc chimera (2019-nCoV Spike Protein RBD, Fc Tag, purchased from Sinobiological) at 2 µg/mL in carbo-free buffer were added to the wells.

    Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay

    Binding of SARS-CoV-2 spike protein to human Siglecs. Human Siglecs ELISA results: wells were coated with the viral spike protein and the binding to different Siglces was evaluated. The experiment has been performed three times in duplicate with similar results, and data were normalized over signal from blocking buffer-coated wells. Error bars indicate standard deviations. OD: Optical density.

    Journal: bioRxiv

    Article Title: Novel ACE2-Independent Carbohydrate-Binding of SARS-CoV-2 Spike Protein to Host Lectins and Lung Microbiota

    doi: 10.1101/2020.05.13.092478

    Figure Lengend Snippet: Binding of SARS-CoV-2 spike protein to human Siglecs. Human Siglecs ELISA results: wells were coated with the viral spike protein and the binding to different Siglces was evaluated. The experiment has been performed three times in duplicate with similar results, and data were normalized over signal from blocking buffer-coated wells. Error bars indicate standard deviations. OD: Optical density.

    Article Snippet: The blocking solution was discarded and 50µL of SARS-CoV-2 spike protein mice-Fc chimera (2019-nCoV Spike Protein RBD, Fc Tag, purchased from Sinobiological) at 2 µg/mL in carbo-free buffer were added to the wells.

    Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Blocking Assay

    Scheme describing the assays employed in this paper. (A) The recombinant protein constructs ACE2-His-Avi (ACE2-Avi) (Acro Biosystems) and SARS-CoV-2 spike protein receptor binding domain-Fc (RBD-Fc) (Sino Biological) were used to model ACE2–RBD binding. (B) AlphaLISA assay system used to monitor ACE2–RBD interacts. Streptavidin donor beads recognize the Avi tag on ACE2. The protein A acceptor beads recognize the Fc tag on RBD. When in proximity the donor beads can be excited with light at 680 nm. This generates singlet oxygen which diffuses to the acceptor beads, causing the acceptor beads to luminesce at 615 nm. (C) The TruHits counterscreen uses streptavidindonor beads which directly interact with biotin acceptor beads. Because no intermediary molecule is needed to bring the donor and acceptor beads in proximity, the TruHits assay can identify compounds which directly interfere with the AlphaLISA readout.

    Journal: ACS Pharmacology & Translational Science

    Article Title: Targeting ACE2–RBD Interaction as a Platform for COVID-19 Therapeutics: Development and Drug-Repurposing Screen of an AlphaLISA Proximity Assay

    doi: 10.1021/acsptsci.0c00161

    Figure Lengend Snippet: Scheme describing the assays employed in this paper. (A) The recombinant protein constructs ACE2-His-Avi (ACE2-Avi) (Acro Biosystems) and SARS-CoV-2 spike protein receptor binding domain-Fc (RBD-Fc) (Sino Biological) were used to model ACE2–RBD binding. (B) AlphaLISA assay system used to monitor ACE2–RBD interacts. Streptavidin donor beads recognize the Avi tag on ACE2. The protein A acceptor beads recognize the Fc tag on RBD. When in proximity the donor beads can be excited with light at 680 nm. This generates singlet oxygen which diffuses to the acceptor beads, causing the acceptor beads to luminesce at 615 nm. (C) The TruHits counterscreen uses streptavidindonor beads which directly interact with biotin acceptor beads. Because no intermediary molecule is needed to bring the donor and acceptor beads in proximity, the TruHits assay can identify compounds which directly interfere with the AlphaLISA readout.

    Article Snippet: RBD-Fc (SARS-CoV-2 spike protein residues 319–541, C-terminal Fc tag; catalog no.40592-V02H), ACE2-His (ACE2 residues 1–740, C-terminal His tag; catalog no. 10108-H08H), and S1-His (spike protein residues 1–667, C-terminal His tag; catalog no. 40150-V08B1) were acquired from Sino Biological (Wayne, PA).

    Techniques: Recombinant, Construct, Binding Assay