sars cov 2 2019 ncov spike s2 antibody rabbit pab  (Sino Biological)


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    Name:
    SARS CoV 2 2019 nCoV Spike S2 Antibody Rabbit PAb
    Description:
    Produced in rabbits immunized with recombinant SARS CoV 2 2019 nCoV Spike S2 Protein Catalog 40590 V08B YP 009724390 1 Ser686 Pro1213 The specific IgG was purified by SARS CoV 2 2019 nCoV Spike S2 affinity chromatography
    Catalog Number:
    40590-T62
    Price:
    None
    Category:
    Primary Antibody
    Reactivity:
    2019 nCoV
    Applications:
    WB,ELISA
    Immunogen:
    Recombinant SARS-CoV-2 / 2019-nCoV Spike/S2 Protein (Catalog#40590-V08B)
    Product Aliases:
    Anti-coronavirus spike Antibody, Anti-cov spike Antibody, Anti-ncov RBD Antibody, Anti-ncov s1 Antibody, Anti-ncov s2 Antibody, Anti-ncov spike Antibody, Anti-NCP-CoV RBD Antibody, Anti-NCP-CoV s1 Antibody, Anti-NCP-CoV s2 Antibody, Anti-NCP-CoV Spike Antibody, Anti-novel coronavirus RBD Antibody, Anti-novel coronavirus s1 Antibody, Anti-novel coronavirus s2 Antibody, Anti-novel coronavirus spike Antibody, Anti-RBD Antibody, Anti-S1 Antibody, Anti-S2 Antibody, Anti-Spike RBD Antibody
    Antibody Type:
    PAb
    Host:
    Rabbit
    Isotype:
    Rabbit IgG
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    Structured Review

    Sino Biological sars cov 2 2019 ncov spike s2 antibody rabbit pab
    Impact of dec-RVKR-CMK on MLVpps. + dec-RVKR-CMK refers to MLVpp produced in HEK293T treated with 75 μM dec-RVKR-CMK at the time of transfection. + furin refers to particles treated with 6 U of recombinant furin for 3 h at 37 °C. (A, B) ± dec-RVKR-CMK and ± furin <t>SARS-CoV-2pp</t> were used to infect Vero E6 and Calu-3 cells. Infectivity was normalized to the – dec-RVKR-CMK, – furin condition in each cell line. Note the change in y -axis scale between A and B. Error bars represent the standard error measurements of three biological replicates ( n = 3). Statistical analysis was performed using an unpaired student’s t test. Not shown: there is a ns difference between the −dec-RVKR-CMK, ± furin conditions (black bars). *, P ≤ 0.05; **, P ≤ 0.01; ns, nonsignificant, P > 0.05. (C) Western blot analysis of SARS-CoV-2pp produced in ± dec-RVKR-CMK and treated with ± furin for S protein content to complement the infection conditions. S was detected using a rabbit antibody against the <t>SARS-CoV-2</t> S2 region. MLV content was detected using a mouse antibody against MLV p30. Band intensities were normalized to the MLV p30 band intensity.
    Produced in rabbits immunized with recombinant SARS CoV 2 2019 nCoV Spike S2 Protein Catalog 40590 V08B YP 009724390 1 Ser686 Pro1213 The specific IgG was purified by SARS CoV 2 2019 nCoV Spike S2 affinity chromatography
    https://www.bioz.com/result/sars cov 2 2019 ncov spike s2 antibody rabbit pab/product/Sino Biological
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sars cov 2 2019 ncov spike s2 antibody rabbit pab - by Bioz Stars, 2021-05
    97/100 stars

    Images

    1) Product Images from "Proteolytic Activation of SARS-CoV-2 Spike at the S1/S2 Boundary: Potential Role of Proteases beyond Furin"

    Article Title: Proteolytic Activation of SARS-CoV-2 Spike at the S1/S2 Boundary: Potential Role of Proteases beyond Furin

    Journal: ACS Infectious Diseases

    doi: 10.1021/acsinfecdis.0c00701

    Impact of dec-RVKR-CMK on MLVpps. + dec-RVKR-CMK refers to MLVpp produced in HEK293T treated with 75 μM dec-RVKR-CMK at the time of transfection. + furin refers to particles treated with 6 U of recombinant furin for 3 h at 37 °C. (A, B) ± dec-RVKR-CMK and ± furin SARS-CoV-2pp were used to infect Vero E6 and Calu-3 cells. Infectivity was normalized to the – dec-RVKR-CMK, – furin condition in each cell line. Note the change in y -axis scale between A and B. Error bars represent the standard error measurements of three biological replicates ( n = 3). Statistical analysis was performed using an unpaired student’s t test. Not shown: there is a ns difference between the −dec-RVKR-CMK, ± furin conditions (black bars). *, P ≤ 0.05; **, P ≤ 0.01; ns, nonsignificant, P > 0.05. (C) Western blot analysis of SARS-CoV-2pp produced in ± dec-RVKR-CMK and treated with ± furin for S protein content to complement the infection conditions. S was detected using a rabbit antibody against the SARS-CoV-2 S2 region. MLV content was detected using a mouse antibody against MLV p30. Band intensities were normalized to the MLV p30 band intensity.
    Figure Legend Snippet: Impact of dec-RVKR-CMK on MLVpps. + dec-RVKR-CMK refers to MLVpp produced in HEK293T treated with 75 μM dec-RVKR-CMK at the time of transfection. + furin refers to particles treated with 6 U of recombinant furin for 3 h at 37 °C. (A, B) ± dec-RVKR-CMK and ± furin SARS-CoV-2pp were used to infect Vero E6 and Calu-3 cells. Infectivity was normalized to the – dec-RVKR-CMK, – furin condition in each cell line. Note the change in y -axis scale between A and B. Error bars represent the standard error measurements of three biological replicates ( n = 3). Statistical analysis was performed using an unpaired student’s t test. Not shown: there is a ns difference between the −dec-RVKR-CMK, ± furin conditions (black bars). *, P ≤ 0.05; **, P ≤ 0.01; ns, nonsignificant, P > 0.05. (C) Western blot analysis of SARS-CoV-2pp produced in ± dec-RVKR-CMK and treated with ± furin for S protein content to complement the infection conditions. S was detected using a rabbit antibody against the SARS-CoV-2 S2 region. MLV content was detected using a mouse antibody against MLV p30. Band intensities were normalized to the MLV p30 band intensity.

    Techniques Used: Produced, Transfection, Recombinant, Infection, Western Blot

    Use of specific furin (alpha1-PDX) and broad furin (dec-RVKR-CMK) inhibitors to investigate furin specificity. (A) Infectivity of particles produced in the presence of broad (dec-RVKR-CMK) and specific (alpha1-PDX) furin inhibitors. The inhibitor was applied at the indicated concentration to producer HEK293T cells. Particles were used to infect Vero E6 cells. Infectivity was normalized to the – condition. Error bars represent the standard error measurements of four biological replicates ( n = 4). Statistical analysis was performed using an unpaired student’s t test compared to the – condition. *, P ≤ 0.05; ns, nonsignificant, P > 0.05. (B) Proteolytic cleavage assay of SARS-CoV-2 S1/S2 peptides to confirm alpha1-PDX and dec-RVKR-CMK inhibition specificity. Alpha1-PDX (2 μM), dec-RVKR-CMK (75 μM), or no inhibitor was incubated with either furin or trypsin recombinant protease and a fluorogenic peptide mimicking the SARS-CoV-2 S1/S2 site (TNSPRRARSVA). Results represent averages, and error bars represent the standard error measurement of three biological replicates ( n = 3).
    Figure Legend Snippet: Use of specific furin (alpha1-PDX) and broad furin (dec-RVKR-CMK) inhibitors to investigate furin specificity. (A) Infectivity of particles produced in the presence of broad (dec-RVKR-CMK) and specific (alpha1-PDX) furin inhibitors. The inhibitor was applied at the indicated concentration to producer HEK293T cells. Particles were used to infect Vero E6 cells. Infectivity was normalized to the – condition. Error bars represent the standard error measurements of four biological replicates ( n = 4). Statistical analysis was performed using an unpaired student’s t test compared to the – condition. *, P ≤ 0.05; ns, nonsignificant, P > 0.05. (B) Proteolytic cleavage assay of SARS-CoV-2 S1/S2 peptides to confirm alpha1-PDX and dec-RVKR-CMK inhibition specificity. Alpha1-PDX (2 μM), dec-RVKR-CMK (75 μM), or no inhibitor was incubated with either furin or trypsin recombinant protease and a fluorogenic peptide mimicking the SARS-CoV-2 S1/S2 site (TNSPRRARSVA). Results represent averages, and error bars represent the standard error measurement of three biological replicates ( n = 3).

    Techniques Used: Infection, Produced, Concentration Assay, Cleavage Assay, Inhibition, Incubation, Recombinant

    Furin cleavage score analysis of CoV S1/S2 cleavage sites. CoV S sequences were analyzed using the ProP 35 1.0 and PiTou 34 3.0 furin prediction algorithm. Bold scores indicate the S1/S2 sequence is predicted to be cleaved by furin. Red box highlights SARS-CoV and SARS-CoV-2 scores for reader’s convenience. Purple lines denote the position of the predicted S1/S2 cleavage site. Basic arginine (R) and lysine (K) residues are highlighted in blue. *For Bat-RmYN02, the sequence number was determined from the S alignment with SARS-CoV-2 S using Geneious. Sequences corresponding to the S1/S2 region of HCoV-HKU1 (AAT98580.1), SARS-CoV (AAT74874.1), SARS-CoV-2 (QHD43416.1), Bat-CoVRaTG13 (QHR63300.2), Bat-SL-CoVZC45 (AVP78031.1), Bat-SL-CoVZXC21 (AVP78042.1), MERS-CoV (AFS88936.1), BatCoV-HKU4 (YP_001039953.1), BatCoV-HKU5 (YP_001039962.1), BatCoV-PML (KC869678), Bat-CoV-HKU9 (YP_001039971), and Influenza A/Chicken/Hong Kong/822.1/01/H5N1 (AF509026.2) were obtained from GenBank. The sequence corresponding to the S1/S2 region of RmYN02 (EPI_ISL_412977) was obtained from GISAID.
    Figure Legend Snippet: Furin cleavage score analysis of CoV S1/S2 cleavage sites. CoV S sequences were analyzed using the ProP 35 1.0 and PiTou 34 3.0 furin prediction algorithm. Bold scores indicate the S1/S2 sequence is predicted to be cleaved by furin. Red box highlights SARS-CoV and SARS-CoV-2 scores for reader’s convenience. Purple lines denote the position of the predicted S1/S2 cleavage site. Basic arginine (R) and lysine (K) residues are highlighted in blue. *For Bat-RmYN02, the sequence number was determined from the S alignment with SARS-CoV-2 S using Geneious. Sequences corresponding to the S1/S2 region of HCoV-HKU1 (AAT98580.1), SARS-CoV (AAT74874.1), SARS-CoV-2 (QHD43416.1), Bat-CoVRaTG13 (QHR63300.2), Bat-SL-CoVZC45 (AVP78031.1), Bat-SL-CoVZXC21 (AVP78042.1), MERS-CoV (AFS88936.1), BatCoV-HKU4 (YP_001039953.1), BatCoV-HKU5 (YP_001039962.1), BatCoV-PML (KC869678), Bat-CoV-HKU9 (YP_001039971), and Influenza A/Chicken/Hong Kong/822.1/01/H5N1 (AF509026.2) were obtained from GenBank. The sequence corresponding to the S1/S2 region of RmYN02 (EPI_ISL_412977) was obtained from GISAID.

    Techniques Used: Sequencing

    Characterization of an MLVpp system. (A, B) MLVpp infectivity in Vero E6 and Calu-3 cells. Cells were infected with MLVpps exhibiting the SARS-CoV-2 S, SARS-CoV S, VSV G, or no envelope protein and assessed for luciferase activity. Error bars represent the standard error measurements of three biological replicates ( n = 3). (C) Western blot analysis of SARS-CoV-2pp and SARS-CoVpp S content. S was detected using a rabbit antibody against the SARS-CoV-2 S2 region and cross reacts against the SARS-CoV S. MLV content was detected using a mouse antibody against MLV p30. The image was cropped from a singular Western blot.
    Figure Legend Snippet: Characterization of an MLVpp system. (A, B) MLVpp infectivity in Vero E6 and Calu-3 cells. Cells were infected with MLVpps exhibiting the SARS-CoV-2 S, SARS-CoV S, VSV G, or no envelope protein and assessed for luciferase activity. Error bars represent the standard error measurements of three biological replicates ( n = 3). (C) Western blot analysis of SARS-CoV-2pp and SARS-CoVpp S content. S was detected using a rabbit antibody against the SARS-CoV-2 S2 region and cross reacts against the SARS-CoV S. MLV content was detected using a mouse antibody against MLV p30. The image was cropped from a singular Western blot.

    Techniques Used: Infection, Luciferase, Activity Assay, Western Blot

    2) Product Images from "Saxifraga spinulosa-Derived Components Rapidly Inactivate Multiple Viruses Including SARS-CoV-2"

    Article Title: Saxifraga spinulosa-Derived Components Rapidly Inactivate Multiple Viruses Including SARS-CoV-2

    Journal: Viruses

    doi: 10.3390/v12070699

    Analysis of the effect of Fr 1C on the SARS-CoV-2 proteins and genome. DMSO and Fr 1C were added to cell culture supernatants containing SARS-CoV-2 and were incubated at 25 °C for 48 h. n = 3 per group. ( A,B ) The images are the results of WB to detect SARS-CoV-2 ( A ) S2 subunit protein and ( B ) NP. ( C ) The image is the result of RT–PCR using NIID_2019-nCoV_N_F2 and R2 primers which amplify 158 bp region on SARS-CoV-2 gene. M: Marker.
    Figure Legend Snippet: Analysis of the effect of Fr 1C on the SARS-CoV-2 proteins and genome. DMSO and Fr 1C were added to cell culture supernatants containing SARS-CoV-2 and were incubated at 25 °C for 48 h. n = 3 per group. ( A,B ) The images are the results of WB to detect SARS-CoV-2 ( A ) S2 subunit protein and ( B ) NP. ( C ) The image is the result of RT–PCR using NIID_2019-nCoV_N_F2 and R2 primers which amplify 158 bp region on SARS-CoV-2 gene. M: Marker.

    Techniques Used: Cell Culture, Incubation, Western Blot, Reverse Transcription Polymerase Chain Reaction, Marker

    3) Product Images from "Proteolytic activation of the SARS-CoV-2 spike S1/S2 site: a re-evaluation of furin cleavage"

    Article Title: Proteolytic activation of the SARS-CoV-2 spike S1/S2 site: a re-evaluation of furin cleavage

    Journal: bioRxiv

    doi: 10.1101/2020.10.04.325522

    Furin cleavage score analysis of CoV S1/S2 cleavage sites. CoV S sequences were analyzed using the ProP 1 1.0 and PiTou 2 3.0 furin prediction algorithm, generating a score with green numbers indicating predicted furin cleavage and red numbers indicating no predicted furin cleavage. Purple lines denote the position of the predicted S1/S2 cleavage site. Basic arginine (R) and lysine (K) residues are highlighted in blue. Sequence numbers refer to position of amino acids within the spike protein. *For Bat-RmYN02, sequence number was determined from S alignment with SARS-CoV-2 S using Geneious. Sequences corresponding to the S1/S2 region of HCoV-HKU1 (AAT98580.1), SARS-CoV (AAT74874.1), SARS-CoV-2 (QHD43416.1), Bat-CoVRaTG13 (QHR63300.2), Bat-SL-CoVZC45 (AVP78031.1), Bat-SL-CoVZXC21 (AVP78042.1), MERS-CoV (AFS88936.1), BatCoV-HKU4 (YP_001039953.1), BatCoV-HKU5 (YP_001039962.1), and Influenza A/Chicken/Hong Kong/822.1/01/H5N1 (AF509026.2) were obtained from GenBank. Sequences corresponding to the S1/S2 and S2’ region of RmYN02 (EPI_ISL_412977) were obtained from GISAID.
    Figure Legend Snippet: Furin cleavage score analysis of CoV S1/S2 cleavage sites. CoV S sequences were analyzed using the ProP 1 1.0 and PiTou 2 3.0 furin prediction algorithm, generating a score with green numbers indicating predicted furin cleavage and red numbers indicating no predicted furin cleavage. Purple lines denote the position of the predicted S1/S2 cleavage site. Basic arginine (R) and lysine (K) residues are highlighted in blue. Sequence numbers refer to position of amino acids within the spike protein. *For Bat-RmYN02, sequence number was determined from S alignment with SARS-CoV-2 S using Geneious. Sequences corresponding to the S1/S2 region of HCoV-HKU1 (AAT98580.1), SARS-CoV (AAT74874.1), SARS-CoV-2 (QHD43416.1), Bat-CoVRaTG13 (QHR63300.2), Bat-SL-CoVZC45 (AVP78031.1), Bat-SL-CoVZXC21 (AVP78042.1), MERS-CoV (AFS88936.1), BatCoV-HKU4 (YP_001039953.1), BatCoV-HKU5 (YP_001039962.1), and Influenza A/Chicken/Hong Kong/822.1/01/H5N1 (AF509026.2) were obtained from GenBank. Sequences corresponding to the S1/S2 and S2’ region of RmYN02 (EPI_ISL_412977) were obtained from GISAID.

    Techniques Used: Sequencing

    Predicted structural model of the SARS-CoV and SARS-CoV-2 S proteins. ( Inset ) Magnification of S1/S2 site with conserved R and S residues (red ribbon) and the unique four amino acid insertion P-R-R-A for SARS-CoV-2 (blue ribbon) are shown. The P’s denote the position of that amino acid from the S1/S2 cleavage site, with P1-P5 referring to amino acids before the cleavage site and P1’ referring to amino acids after the cleavage site.
    Figure Legend Snippet: Predicted structural model of the SARS-CoV and SARS-CoV-2 S proteins. ( Inset ) Magnification of S1/S2 site with conserved R and S residues (red ribbon) and the unique four amino acid insertion P-R-R-A for SARS-CoV-2 (blue ribbon) are shown. The P’s denote the position of that amino acid from the S1/S2 cleavage site, with P1-P5 referring to amino acids before the cleavage site and P1’ referring to amino acids after the cleavage site.

    Techniques Used:

    Characterization of MLVpp system. ( A and B ) MLVpp infectivity in Vero E6 and Calu-3 cells. Cells were infected with MLVpps exhibiting the SARS-CoV-2 S, SARS-CoV S, VSV G or no envelope protein and assessed for luciferase activity. Error bars represent the standard error measurements of three biological replicates (n=3). ( C ) Western blot analysis of SARS-2pp and SARSpp S content. S was detected using a rabbit antibody against the SARS-CoV-2 S2 region that cross reacts against the SARS-CoV S. MLV content was detected using a mouse antibody against MLVp30. Image cropped from a singular Western blot and lanes are reshown in Figure 3C and 5C .
    Figure Legend Snippet: Characterization of MLVpp system. ( A and B ) MLVpp infectivity in Vero E6 and Calu-3 cells. Cells were infected with MLVpps exhibiting the SARS-CoV-2 S, SARS-CoV S, VSV G or no envelope protein and assessed for luciferase activity. Error bars represent the standard error measurements of three biological replicates (n=3). ( C ) Western blot analysis of SARS-2pp and SARSpp S content. S was detected using a rabbit antibody against the SARS-CoV-2 S2 region that cross reacts against the SARS-CoV S. MLV content was detected using a mouse antibody against MLVp30. Image cropped from a singular Western blot and lanes are reshown in Figure 3C and 5C .

    Techniques Used: Infection, Luciferase, Activity Assay, Western Blot

    4) Product Images from "Difference in levels of SARS-CoV-2 S1 and S2 subunits- and nucleocapsid protein-reactive SIgM/IgM, IgG and SIgA/IgA antibodies in human milk"

    Article Title: Difference in levels of SARS-CoV-2 S1 and S2 subunits- and nucleocapsid protein-reactive SIgM/IgM, IgG and SIgA/IgA antibodies in human milk

    Journal: Journal of Perinatology

    doi: 10.1038/s41372-020-00805-w

    The levels of SARS-CoV-2-reactive to S1 and S2 subunits (S1 + S2) and nucleocapsid in human milk collected during the COVID-19 pandemic. The levels of a S1 + S2- and b nucleocapsid-reactive secretory IgM (SIgM)/IgM, IgG, and secretory IgA (SIgA)/IgA in human milk. Values are mean ± SD, n = 41 for women. Asterisks show statistically significant differences between variables (*** p
    Figure Legend Snippet: The levels of SARS-CoV-2-reactive to S1 and S2 subunits (S1 + S2) and nucleocapsid in human milk collected during the COVID-19 pandemic. The levels of a S1 + S2- and b nucleocapsid-reactive secretory IgM (SIgM)/IgM, IgG, and secretory IgA (SIgA)/IgA in human milk. Values are mean ± SD, n = 41 for women. Asterisks show statistically significant differences between variables (*** p

    Techniques Used:

    Regression linear between antibodies reactive to SARS-CoV-2 S1 and S2 subunits (S1 + S2) and nucleocapsid in human milk collected during the COVID-19 pandemic in 41 women. a Positive correlation of S1 + S2-reactive secretory IgA (SIgA)/IgA and S1 + S2-reactive secretory IgM (SIgM)/IgM. b Positive correlation between S1 + S2-reactive SIgA/IgA and nucleocapsid-reactive SIgA/IgA. c Positive correlation between nucleocapsid-reactive SIgA/IgA and SIgM/IgM. d Positive correlation between nucleocapsid-reactive SIgM/IgM and S1 + S2-reactive SIgM/IgM. Pearson correlation coefficients ( r ) were determined when p
    Figure Legend Snippet: Regression linear between antibodies reactive to SARS-CoV-2 S1 and S2 subunits (S1 + S2) and nucleocapsid in human milk collected during the COVID-19 pandemic in 41 women. a Positive correlation of S1 + S2-reactive secretory IgA (SIgA)/IgA and S1 + S2-reactive secretory IgM (SIgM)/IgM. b Positive correlation between S1 + S2-reactive SIgA/IgA and nucleocapsid-reactive SIgA/IgA. c Positive correlation between nucleocapsid-reactive SIgA/IgA and SIgM/IgM. d Positive correlation between nucleocapsid-reactive SIgM/IgM and S1 + S2-reactive SIgM/IgM. Pearson correlation coefficients ( r ) were determined when p

    Techniques Used:

    The levels of antibodies reactive to SARS-CoV-2 S1 and S2 subunits (S1 + S2) and nucleocapsid in human milk collected during the COVID-19 pandemic (2020-HM) and 2 years prior this pandemic (2018-HM). Levels of S1 + S2-reactive a secretory IgM (SIgM)/IgM, b IgG and c secretory IgA (SIgA)/IgA in 2020-HM and 2018-HM. Levels of nucleocapsid-reactive d SIgM/IgM, e IgG and f SIgA/IgA in 2020-HM and 2018-HM. Values are mean ± SD, n = 41 for 2020-HM and n = 16 for 2018-HM. Asterisks show statistically significant differences between variables (* p
    Figure Legend Snippet: The levels of antibodies reactive to SARS-CoV-2 S1 and S2 subunits (S1 + S2) and nucleocapsid in human milk collected during the COVID-19 pandemic (2020-HM) and 2 years prior this pandemic (2018-HM). Levels of S1 + S2-reactive a secretory IgM (SIgM)/IgM, b IgG and c secretory IgA (SIgA)/IgA in 2020-HM and 2018-HM. Levels of nucleocapsid-reactive d SIgM/IgM, e IgG and f SIgA/IgA in 2020-HM and 2018-HM. Values are mean ± SD, n = 41 for 2020-HM and n = 16 for 2018-HM. Asterisks show statistically significant differences between variables (* p

    Techniques Used:

    Percentage of detected antibodies reactive to S1 and S2 subunits (S1 + S2), and nucleocapsid from SARS-CoV-2 in human milk collected during the COVID-19 pandemic. S1+S2-reactive a secretory IgM (SIgM)/IgM, b IgG and c secretory IgA (SIgA)/IgA in human milk from 41 women. Nucleocapsid-reactive d SIgM/IgM, e IgG and f SIgA/IgA. Milk expression period was from 30/02/20 to 03/04/20 in the United States.
    Figure Legend Snippet: Percentage of detected antibodies reactive to S1 and S2 subunits (S1 + S2), and nucleocapsid from SARS-CoV-2 in human milk collected during the COVID-19 pandemic. S1+S2-reactive a secretory IgM (SIgM)/IgM, b IgG and c secretory IgA (SIgA)/IgA in human milk from 41 women. Nucleocapsid-reactive d SIgM/IgM, e IgG and f SIgA/IgA. Milk expression period was from 30/02/20 to 03/04/20 in the United States.

    Techniques Used: Expressing

    5) Product Images from "Difference in levels of SARS-CoV-2 S1 and S2 subunits- and nucleocapsid protein-reactive SIgM/IgM, IgG and SIgA/IgA antibodies in human milk"

    Article Title: Difference in levels of SARS-CoV-2 S1 and S2 subunits- and nucleocapsid protein-reactive SIgM/IgM, IgG and SIgA/IgA antibodies in human milk

    Journal: Journal of Perinatology

    doi: 10.1038/s41372-020-00805-w

    The levels of SARS-CoV-2-reactive to S1 and S2 subunits (S1 + S2) and nucleocapsid in human milk collected during the COVID-19 pandemic. The levels of a S1 + S2- and b nucleocapsid-reactive secretory IgM (SIgM)/IgM, IgG, and secretory IgA (SIgA)/IgA in human milk. Values are mean ± SD, n = 41 for women. Asterisks show statistically significant differences between variables (*** p
    Figure Legend Snippet: The levels of SARS-CoV-2-reactive to S1 and S2 subunits (S1 + S2) and nucleocapsid in human milk collected during the COVID-19 pandemic. The levels of a S1 + S2- and b nucleocapsid-reactive secretory IgM (SIgM)/IgM, IgG, and secretory IgA (SIgA)/IgA in human milk. Values are mean ± SD, n = 41 for women. Asterisks show statistically significant differences between variables (*** p

    Techniques Used:

    Regression linear between antibodies reactive to SARS-CoV-2 S1 and S2 subunits (S1 + S2) and nucleocapsid in human milk collected during the COVID-19 pandemic in 41 women. a Positive correlation of S1 + S2-reactive secretory IgA (SIgA)/IgA and S1 + S2-reactive secretory IgM (SIgM)/IgM. b Positive correlation between S1 + S2-reactive SIgA/IgA and nucleocapsid-reactive SIgA/IgA. c Positive correlation between nucleocapsid-reactive SIgA/IgA and SIgM/IgM. d Positive correlation between nucleocapsid-reactive SIgM/IgM and S1 + S2-reactive SIgM/IgM. Pearson correlation coefficients ( r ) were determined when p
    Figure Legend Snippet: Regression linear between antibodies reactive to SARS-CoV-2 S1 and S2 subunits (S1 + S2) and nucleocapsid in human milk collected during the COVID-19 pandemic in 41 women. a Positive correlation of S1 + S2-reactive secretory IgA (SIgA)/IgA and S1 + S2-reactive secretory IgM (SIgM)/IgM. b Positive correlation between S1 + S2-reactive SIgA/IgA and nucleocapsid-reactive SIgA/IgA. c Positive correlation between nucleocapsid-reactive SIgA/IgA and SIgM/IgM. d Positive correlation between nucleocapsid-reactive SIgM/IgM and S1 + S2-reactive SIgM/IgM. Pearson correlation coefficients ( r ) were determined when p

    Techniques Used:

    The levels of antibodies reactive to SARS-CoV-2 S1 and S2 subunits (S1 + S2) and nucleocapsid in human milk collected during the COVID-19 pandemic (2020-HM) and 2 years prior this pandemic (2018-HM). Levels of S1 + S2-reactive a secretory IgM (SIgM)/IgM, b IgG and c secretory IgA (SIgA)/IgA in 2020-HM and 2018-HM. Levels of nucleocapsid-reactive d SIgM/IgM, e IgG and f SIgA/IgA in 2020-HM and 2018-HM. Values are mean ± SD, n = 41 for 2020-HM and n = 16 for 2018-HM. Asterisks show statistically significant differences between variables (* p
    Figure Legend Snippet: The levels of antibodies reactive to SARS-CoV-2 S1 and S2 subunits (S1 + S2) and nucleocapsid in human milk collected during the COVID-19 pandemic (2020-HM) and 2 years prior this pandemic (2018-HM). Levels of S1 + S2-reactive a secretory IgM (SIgM)/IgM, b IgG and c secretory IgA (SIgA)/IgA in 2020-HM and 2018-HM. Levels of nucleocapsid-reactive d SIgM/IgM, e IgG and f SIgA/IgA in 2020-HM and 2018-HM. Values are mean ± SD, n = 41 for 2020-HM and n = 16 for 2018-HM. Asterisks show statistically significant differences between variables (* p

    Techniques Used:

    Percentage of detected antibodies reactive to S1 and S2 subunits (S1 + S2), and nucleocapsid from SARS-CoV-2 in human milk collected during the COVID-19 pandemic. S1+S2-reactive a secretory IgM (SIgM)/IgM, b IgG and c secretory IgA (SIgA)/IgA in human milk from 41 women. Nucleocapsid-reactive d SIgM/IgM, e IgG and f SIgA/IgA. Milk expression period was from 30/02/20 to 03/04/20 in the United States.
    Figure Legend Snippet: Percentage of detected antibodies reactive to S1 and S2 subunits (S1 + S2), and nucleocapsid from SARS-CoV-2 in human milk collected during the COVID-19 pandemic. S1+S2-reactive a secretory IgM (SIgM)/IgM, b IgG and c secretory IgA (SIgA)/IgA in human milk from 41 women. Nucleocapsid-reactive d SIgM/IgM, e IgG and f SIgA/IgA. Milk expression period was from 30/02/20 to 03/04/20 in the United States.

    Techniques Used: Expressing

    6) Product Images from "Proteolytic Activation of SARS-CoV-2 Spike at the S1/S2 Boundary: Potential Role of Proteases beyond Furin"

    Article Title: Proteolytic Activation of SARS-CoV-2 Spike at the S1/S2 Boundary: Potential Role of Proteases beyond Furin

    Journal: ACS Infectious Diseases

    doi: 10.1021/acsinfecdis.0c00701

    Impact of dec-RVKR-CMK on MLVpps. + dec-RVKR-CMK refers to MLVpp produced in HEK293T treated with 75 μM dec-RVKR-CMK at the time of transfection. + furin refers to particles treated with 6 U of recombinant furin for 3 h at 37 °C. (A, B) ± dec-RVKR-CMK and ± furin SARS-CoV-2pp were used to infect Vero E6 and Calu-3 cells. Infectivity was normalized to the – dec-RVKR-CMK, – furin condition in each cell line. Note the change in y -axis scale between A and B. Error bars represent the standard error measurements of three biological replicates ( n = 3). Statistical analysis was performed using an unpaired student’s t test. Not shown: there is a ns difference between the −dec-RVKR-CMK, ± furin conditions (black bars). *, P ≤ 0.05; **, P ≤ 0.01; ns, nonsignificant, P > 0.05. (C) Western blot analysis of SARS-CoV-2pp produced in ± dec-RVKR-CMK and treated with ± furin for S protein content to complement the infection conditions. S was detected using a rabbit antibody against the SARS-CoV-2 S2 region. MLV content was detected using a mouse antibody against MLV p30. Band intensities were normalized to the MLV p30 band intensity.
    Figure Legend Snippet: Impact of dec-RVKR-CMK on MLVpps. + dec-RVKR-CMK refers to MLVpp produced in HEK293T treated with 75 μM dec-RVKR-CMK at the time of transfection. + furin refers to particles treated with 6 U of recombinant furin for 3 h at 37 °C. (A, B) ± dec-RVKR-CMK and ± furin SARS-CoV-2pp were used to infect Vero E6 and Calu-3 cells. Infectivity was normalized to the – dec-RVKR-CMK, – furin condition in each cell line. Note the change in y -axis scale between A and B. Error bars represent the standard error measurements of three biological replicates ( n = 3). Statistical analysis was performed using an unpaired student’s t test. Not shown: there is a ns difference between the −dec-RVKR-CMK, ± furin conditions (black bars). *, P ≤ 0.05; **, P ≤ 0.01; ns, nonsignificant, P > 0.05. (C) Western blot analysis of SARS-CoV-2pp produced in ± dec-RVKR-CMK and treated with ± furin for S protein content to complement the infection conditions. S was detected using a rabbit antibody against the SARS-CoV-2 S2 region. MLV content was detected using a mouse antibody against MLV p30. Band intensities were normalized to the MLV p30 band intensity.

    Techniques Used: Produced, Transfection, Recombinant, Infection, Western Blot

    Use of specific furin (alpha1-PDX) and broad furin (dec-RVKR-CMK) inhibitors to investigate furin specificity. (A) Infectivity of particles produced in the presence of broad (dec-RVKR-CMK) and specific (alpha1-PDX) furin inhibitors. The inhibitor was applied at the indicated concentration to producer HEK293T cells. Particles were used to infect Vero E6 cells. Infectivity was normalized to the – condition. Error bars represent the standard error measurements of four biological replicates ( n = 4). Statistical analysis was performed using an unpaired student’s t test compared to the – condition. *, P ≤ 0.05; ns, nonsignificant, P > 0.05. (B) Proteolytic cleavage assay of SARS-CoV-2 S1/S2 peptides to confirm alpha1-PDX and dec-RVKR-CMK inhibition specificity. Alpha1-PDX (2 μM), dec-RVKR-CMK (75 μM), or no inhibitor was incubated with either furin or trypsin recombinant protease and a fluorogenic peptide mimicking the SARS-CoV-2 S1/S2 site (TNSPRRARSVA). Results represent averages, and error bars represent the standard error measurement of three biological replicates ( n = 3).
    Figure Legend Snippet: Use of specific furin (alpha1-PDX) and broad furin (dec-RVKR-CMK) inhibitors to investigate furin specificity. (A) Infectivity of particles produced in the presence of broad (dec-RVKR-CMK) and specific (alpha1-PDX) furin inhibitors. The inhibitor was applied at the indicated concentration to producer HEK293T cells. Particles were used to infect Vero E6 cells. Infectivity was normalized to the – condition. Error bars represent the standard error measurements of four biological replicates ( n = 4). Statistical analysis was performed using an unpaired student’s t test compared to the – condition. *, P ≤ 0.05; ns, nonsignificant, P > 0.05. (B) Proteolytic cleavage assay of SARS-CoV-2 S1/S2 peptides to confirm alpha1-PDX and dec-RVKR-CMK inhibition specificity. Alpha1-PDX (2 μM), dec-RVKR-CMK (75 μM), or no inhibitor was incubated with either furin or trypsin recombinant protease and a fluorogenic peptide mimicking the SARS-CoV-2 S1/S2 site (TNSPRRARSVA). Results represent averages, and error bars represent the standard error measurement of three biological replicates ( n = 3).

    Techniques Used: Infection, Produced, Concentration Assay, Cleavage Assay, Inhibition, Incubation, Recombinant

    Furin cleavage score analysis of CoV S1/S2 cleavage sites. CoV S sequences were analyzed using the ProP 35 1.0 and PiTou 34 3.0 furin prediction algorithm. Bold scores indicate the S1/S2 sequence is predicted to be cleaved by furin. Red box highlights SARS-CoV and SARS-CoV-2 scores for reader’s convenience. Purple lines denote the position of the predicted S1/S2 cleavage site. Basic arginine (R) and lysine (K) residues are highlighted in blue. *For Bat-RmYN02, the sequence number was determined from the S alignment with SARS-CoV-2 S using Geneious. Sequences corresponding to the S1/S2 region of HCoV-HKU1 (AAT98580.1), SARS-CoV (AAT74874.1), SARS-CoV-2 (QHD43416.1), Bat-CoVRaTG13 (QHR63300.2), Bat-SL-CoVZC45 (AVP78031.1), Bat-SL-CoVZXC21 (AVP78042.1), MERS-CoV (AFS88936.1), BatCoV-HKU4 (YP_001039953.1), BatCoV-HKU5 (YP_001039962.1), BatCoV-PML (KC869678), Bat-CoV-HKU9 (YP_001039971), and Influenza A/Chicken/Hong Kong/822.1/01/H5N1 (AF509026.2) were obtained from GenBank. The sequence corresponding to the S1/S2 region of RmYN02 (EPI_ISL_412977) was obtained from GISAID.
    Figure Legend Snippet: Furin cleavage score analysis of CoV S1/S2 cleavage sites. CoV S sequences were analyzed using the ProP 35 1.0 and PiTou 34 3.0 furin prediction algorithm. Bold scores indicate the S1/S2 sequence is predicted to be cleaved by furin. Red box highlights SARS-CoV and SARS-CoV-2 scores for reader’s convenience. Purple lines denote the position of the predicted S1/S2 cleavage site. Basic arginine (R) and lysine (K) residues are highlighted in blue. *For Bat-RmYN02, the sequence number was determined from the S alignment with SARS-CoV-2 S using Geneious. Sequences corresponding to the S1/S2 region of HCoV-HKU1 (AAT98580.1), SARS-CoV (AAT74874.1), SARS-CoV-2 (QHD43416.1), Bat-CoVRaTG13 (QHR63300.2), Bat-SL-CoVZC45 (AVP78031.1), Bat-SL-CoVZXC21 (AVP78042.1), MERS-CoV (AFS88936.1), BatCoV-HKU4 (YP_001039953.1), BatCoV-HKU5 (YP_001039962.1), BatCoV-PML (KC869678), Bat-CoV-HKU9 (YP_001039971), and Influenza A/Chicken/Hong Kong/822.1/01/H5N1 (AF509026.2) were obtained from GenBank. The sequence corresponding to the S1/S2 region of RmYN02 (EPI_ISL_412977) was obtained from GISAID.

    Techniques Used: Sequencing

    Characterization of an MLVpp system. (A, B) MLVpp infectivity in Vero E6 and Calu-3 cells. Cells were infected with MLVpps exhibiting the SARS-CoV-2 S, SARS-CoV S, VSV G, or no envelope protein and assessed for luciferase activity. Error bars represent the standard error measurements of three biological replicates ( n = 3). (C) Western blot analysis of SARS-CoV-2pp and SARS-CoVpp S content. S was detected using a rabbit antibody against the SARS-CoV-2 S2 region and cross reacts against the SARS-CoV S. MLV content was detected using a mouse antibody against MLV p30. The image was cropped from a singular Western blot.
    Figure Legend Snippet: Characterization of an MLVpp system. (A, B) MLVpp infectivity in Vero E6 and Calu-3 cells. Cells were infected with MLVpps exhibiting the SARS-CoV-2 S, SARS-CoV S, VSV G, or no envelope protein and assessed for luciferase activity. Error bars represent the standard error measurements of three biological replicates ( n = 3). (C) Western blot analysis of SARS-CoV-2pp and SARS-CoVpp S content. S was detected using a rabbit antibody against the SARS-CoV-2 S2 region and cross reacts against the SARS-CoV S. MLV content was detected using a mouse antibody against MLV p30. The image was cropped from a singular Western blot.

    Techniques Used: Infection, Luciferase, Activity Assay, Western Blot

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    Article Title: Difference in levels of SARS-CoV-2 S1 and S2 subunits- and nucleocapsid protein-reactive SIgM/IgM, IgG and SIgA/IgA antibodies in human milk
    Article Snippet: SARS-CoV-2 S1 + S2- and nucleocapsid-reactive antibodies The levels of SARS-CoV-2 S1 + S2 and nucleocapsid-reactive SIgM/IgM, IgG and SIgA/IgA were determined using ELISAs that were adapted from our previous publications [ , – ].

    Article Title: The SARS-CoV-2 envelope and membrane proteins modulate maturation and retention of the spike protein, allowing assembly of virus-like particles
    Article Snippet: AntibodiesMouse anti-actin (clone AC74, Sigma-Aldrich), rabbit anti-SARS-CoV2 S2, mouse anti-SARS-CoV2 S1 and mouse anti-SARS-CoV2 N (Sino Biological), mouse anti-GFP (Roche), anti-VSV-G (41A1), and rabbit anti-GM130 (clone EP892Y, Abcam) were used according to the providers’ instructions.

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    Sino Biological sars cov 2 2019 ncov spike s2 antibody rabbit pab
    Impact of dec-RVKR-CMK on MLVpps. + dec-RVKR-CMK refers to MLVpp produced in HEK293T treated with 75 μM dec-RVKR-CMK at the time of transfection. + furin refers to particles treated with 6 U of recombinant furin for 3 h at 37 °C. (A, B) ± dec-RVKR-CMK and ± furin <t>SARS-CoV-2pp</t> were used to infect Vero E6 and Calu-3 cells. Infectivity was normalized to the – dec-RVKR-CMK, – furin condition in each cell line. Note the change in y -axis scale between A and B. Error bars represent the standard error measurements of three biological replicates ( n = 3). Statistical analysis was performed using an unpaired student’s t test. Not shown: there is a ns difference between the −dec-RVKR-CMK, ± furin conditions (black bars). *, P ≤ 0.05; **, P ≤ 0.01; ns, nonsignificant, P > 0.05. (C) Western blot analysis of SARS-CoV-2pp produced in ± dec-RVKR-CMK and treated with ± furin for S protein content to complement the infection conditions. S was detected using a rabbit antibody against the <t>SARS-CoV-2</t> S2 region. MLV content was detected using a mouse antibody against MLV p30. Band intensities were normalized to the MLV p30 band intensity.
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    Impact of dec-RVKR-CMK on MLVpps. + dec-RVKR-CMK refers to MLVpp produced in HEK293T treated with 75 μM dec-RVKR-CMK at the time of transfection. + furin refers to particles treated with 6 U of recombinant furin for 3 h at 37 °C. (A, B) ± dec-RVKR-CMK and ± furin SARS-CoV-2pp were used to infect Vero E6 and Calu-3 cells. Infectivity was normalized to the – dec-RVKR-CMK, – furin condition in each cell line. Note the change in y -axis scale between A and B. Error bars represent the standard error measurements of three biological replicates ( n = 3). Statistical analysis was performed using an unpaired student’s t test. Not shown: there is a ns difference between the −dec-RVKR-CMK, ± furin conditions (black bars). *, P ≤ 0.05; **, P ≤ 0.01; ns, nonsignificant, P > 0.05. (C) Western blot analysis of SARS-CoV-2pp produced in ± dec-RVKR-CMK and treated with ± furin for S protein content to complement the infection conditions. S was detected using a rabbit antibody against the SARS-CoV-2 S2 region. MLV content was detected using a mouse antibody against MLV p30. Band intensities were normalized to the MLV p30 band intensity.

    Journal: ACS Infectious Diseases

    Article Title: Proteolytic Activation of SARS-CoV-2 Spike at the S1/S2 Boundary: Potential Role of Proteases beyond Furin

    doi: 10.1021/acsinfecdis.0c00701

    Figure Lengend Snippet: Impact of dec-RVKR-CMK on MLVpps. + dec-RVKR-CMK refers to MLVpp produced in HEK293T treated with 75 μM dec-RVKR-CMK at the time of transfection. + furin refers to particles treated with 6 U of recombinant furin for 3 h at 37 °C. (A, B) ± dec-RVKR-CMK and ± furin SARS-CoV-2pp were used to infect Vero E6 and Calu-3 cells. Infectivity was normalized to the – dec-RVKR-CMK, – furin condition in each cell line. Note the change in y -axis scale between A and B. Error bars represent the standard error measurements of three biological replicates ( n = 3). Statistical analysis was performed using an unpaired student’s t test. Not shown: there is a ns difference between the −dec-RVKR-CMK, ± furin conditions (black bars). *, P ≤ 0.05; **, P ≤ 0.01; ns, nonsignificant, P > 0.05. (C) Western blot analysis of SARS-CoV-2pp produced in ± dec-RVKR-CMK and treated with ± furin for S protein content to complement the infection conditions. S was detected using a rabbit antibody against the SARS-CoV-2 S2 region. MLV content was detected using a mouse antibody against MLV p30. Band intensities were normalized to the MLV p30 band intensity.

    Article Snippet: SARS-CoV-2 and SARS-CoV S were detected using a rabbit polyclonal antibody that recognizes the S2 domain (Cat: 40590-T62, Sinobiological) and an AlexaFluor 488 goat anti-rabbit secondary antibody.

    Techniques: Produced, Transfection, Recombinant, Infection, Western Blot

    Use of specific furin (alpha1-PDX) and broad furin (dec-RVKR-CMK) inhibitors to investigate furin specificity. (A) Infectivity of particles produced in the presence of broad (dec-RVKR-CMK) and specific (alpha1-PDX) furin inhibitors. The inhibitor was applied at the indicated concentration to producer HEK293T cells. Particles were used to infect Vero E6 cells. Infectivity was normalized to the – condition. Error bars represent the standard error measurements of four biological replicates ( n = 4). Statistical analysis was performed using an unpaired student’s t test compared to the – condition. *, P ≤ 0.05; ns, nonsignificant, P > 0.05. (B) Proteolytic cleavage assay of SARS-CoV-2 S1/S2 peptides to confirm alpha1-PDX and dec-RVKR-CMK inhibition specificity. Alpha1-PDX (2 μM), dec-RVKR-CMK (75 μM), or no inhibitor was incubated with either furin or trypsin recombinant protease and a fluorogenic peptide mimicking the SARS-CoV-2 S1/S2 site (TNSPRRARSVA). Results represent averages, and error bars represent the standard error measurement of three biological replicates ( n = 3).

    Journal: ACS Infectious Diseases

    Article Title: Proteolytic Activation of SARS-CoV-2 Spike at the S1/S2 Boundary: Potential Role of Proteases beyond Furin

    doi: 10.1021/acsinfecdis.0c00701

    Figure Lengend Snippet: Use of specific furin (alpha1-PDX) and broad furin (dec-RVKR-CMK) inhibitors to investigate furin specificity. (A) Infectivity of particles produced in the presence of broad (dec-RVKR-CMK) and specific (alpha1-PDX) furin inhibitors. The inhibitor was applied at the indicated concentration to producer HEK293T cells. Particles were used to infect Vero E6 cells. Infectivity was normalized to the – condition. Error bars represent the standard error measurements of four biological replicates ( n = 4). Statistical analysis was performed using an unpaired student’s t test compared to the – condition. *, P ≤ 0.05; ns, nonsignificant, P > 0.05. (B) Proteolytic cleavage assay of SARS-CoV-2 S1/S2 peptides to confirm alpha1-PDX and dec-RVKR-CMK inhibition specificity. Alpha1-PDX (2 μM), dec-RVKR-CMK (75 μM), or no inhibitor was incubated with either furin or trypsin recombinant protease and a fluorogenic peptide mimicking the SARS-CoV-2 S1/S2 site (TNSPRRARSVA). Results represent averages, and error bars represent the standard error measurement of three biological replicates ( n = 3).

    Article Snippet: SARS-CoV-2 and SARS-CoV S were detected using a rabbit polyclonal antibody that recognizes the S2 domain (Cat: 40590-T62, Sinobiological) and an AlexaFluor 488 goat anti-rabbit secondary antibody.

    Techniques: Infection, Produced, Concentration Assay, Cleavage Assay, Inhibition, Incubation, Recombinant

    Furin cleavage score analysis of CoV S1/S2 cleavage sites. CoV S sequences were analyzed using the ProP 35 1.0 and PiTou 34 3.0 furin prediction algorithm. Bold scores indicate the S1/S2 sequence is predicted to be cleaved by furin. Red box highlights SARS-CoV and SARS-CoV-2 scores for reader’s convenience. Purple lines denote the position of the predicted S1/S2 cleavage site. Basic arginine (R) and lysine (K) residues are highlighted in blue. *For Bat-RmYN02, the sequence number was determined from the S alignment with SARS-CoV-2 S using Geneious. Sequences corresponding to the S1/S2 region of HCoV-HKU1 (AAT98580.1), SARS-CoV (AAT74874.1), SARS-CoV-2 (QHD43416.1), Bat-CoVRaTG13 (QHR63300.2), Bat-SL-CoVZC45 (AVP78031.1), Bat-SL-CoVZXC21 (AVP78042.1), MERS-CoV (AFS88936.1), BatCoV-HKU4 (YP_001039953.1), BatCoV-HKU5 (YP_001039962.1), BatCoV-PML (KC869678), Bat-CoV-HKU9 (YP_001039971), and Influenza A/Chicken/Hong Kong/822.1/01/H5N1 (AF509026.2) were obtained from GenBank. The sequence corresponding to the S1/S2 region of RmYN02 (EPI_ISL_412977) was obtained from GISAID.

    Journal: ACS Infectious Diseases

    Article Title: Proteolytic Activation of SARS-CoV-2 Spike at the S1/S2 Boundary: Potential Role of Proteases beyond Furin

    doi: 10.1021/acsinfecdis.0c00701

    Figure Lengend Snippet: Furin cleavage score analysis of CoV S1/S2 cleavage sites. CoV S sequences were analyzed using the ProP 35 1.0 and PiTou 34 3.0 furin prediction algorithm. Bold scores indicate the S1/S2 sequence is predicted to be cleaved by furin. Red box highlights SARS-CoV and SARS-CoV-2 scores for reader’s convenience. Purple lines denote the position of the predicted S1/S2 cleavage site. Basic arginine (R) and lysine (K) residues are highlighted in blue. *For Bat-RmYN02, the sequence number was determined from the S alignment with SARS-CoV-2 S using Geneious. Sequences corresponding to the S1/S2 region of HCoV-HKU1 (AAT98580.1), SARS-CoV (AAT74874.1), SARS-CoV-2 (QHD43416.1), Bat-CoVRaTG13 (QHR63300.2), Bat-SL-CoVZC45 (AVP78031.1), Bat-SL-CoVZXC21 (AVP78042.1), MERS-CoV (AFS88936.1), BatCoV-HKU4 (YP_001039953.1), BatCoV-HKU5 (YP_001039962.1), BatCoV-PML (KC869678), Bat-CoV-HKU9 (YP_001039971), and Influenza A/Chicken/Hong Kong/822.1/01/H5N1 (AF509026.2) were obtained from GenBank. The sequence corresponding to the S1/S2 region of RmYN02 (EPI_ISL_412977) was obtained from GISAID.

    Article Snippet: SARS-CoV-2 and SARS-CoV S were detected using a rabbit polyclonal antibody that recognizes the S2 domain (Cat: 40590-T62, Sinobiological) and an AlexaFluor 488 goat anti-rabbit secondary antibody.

    Techniques: Sequencing

    Characterization of an MLVpp system. (A, B) MLVpp infectivity in Vero E6 and Calu-3 cells. Cells were infected with MLVpps exhibiting the SARS-CoV-2 S, SARS-CoV S, VSV G, or no envelope protein and assessed for luciferase activity. Error bars represent the standard error measurements of three biological replicates ( n = 3). (C) Western blot analysis of SARS-CoV-2pp and SARS-CoVpp S content. S was detected using a rabbit antibody against the SARS-CoV-2 S2 region and cross reacts against the SARS-CoV S. MLV content was detected using a mouse antibody against MLV p30. The image was cropped from a singular Western blot.

    Journal: ACS Infectious Diseases

    Article Title: Proteolytic Activation of SARS-CoV-2 Spike at the S1/S2 Boundary: Potential Role of Proteases beyond Furin

    doi: 10.1021/acsinfecdis.0c00701

    Figure Lengend Snippet: Characterization of an MLVpp system. (A, B) MLVpp infectivity in Vero E6 and Calu-3 cells. Cells were infected with MLVpps exhibiting the SARS-CoV-2 S, SARS-CoV S, VSV G, or no envelope protein and assessed for luciferase activity. Error bars represent the standard error measurements of three biological replicates ( n = 3). (C) Western blot analysis of SARS-CoV-2pp and SARS-CoVpp S content. S was detected using a rabbit antibody against the SARS-CoV-2 S2 region and cross reacts against the SARS-CoV S. MLV content was detected using a mouse antibody against MLV p30. The image was cropped from a singular Western blot.

    Article Snippet: SARS-CoV-2 and SARS-CoV S were detected using a rabbit polyclonal antibody that recognizes the S2 domain (Cat: 40590-T62, Sinobiological) and an AlexaFluor 488 goat anti-rabbit secondary antibody.

    Techniques: Infection, Luciferase, Activity Assay, Western Blot

    Analysis of the effect of Fr 1C on the SARS-CoV-2 proteins and genome. DMSO and Fr 1C were added to cell culture supernatants containing SARS-CoV-2 and were incubated at 25 °C for 48 h. n = 3 per group. ( A,B ) The images are the results of WB to detect SARS-CoV-2 ( A ) S2 subunit protein and ( B ) NP. ( C ) The image is the result of RT–PCR using NIID_2019-nCoV_N_F2 and R2 primers which amplify 158 bp region on SARS-CoV-2 gene. M: Marker.

    Journal: Viruses

    Article Title: Saxifraga spinulosa-Derived Components Rapidly Inactivate Multiple Viruses Including SARS-CoV-2

    doi: 10.3390/v12070699

    Figure Lengend Snippet: Analysis of the effect of Fr 1C on the SARS-CoV-2 proteins and genome. DMSO and Fr 1C were added to cell culture supernatants containing SARS-CoV-2 and were incubated at 25 °C for 48 h. n = 3 per group. ( A,B ) The images are the results of WB to detect SARS-CoV-2 ( A ) S2 subunit protein and ( B ) NP. ( C ) The image is the result of RT–PCR using NIID_2019-nCoV_N_F2 and R2 primers which amplify 158 bp region on SARS-CoV-2 gene. M: Marker.

    Article Snippet: SARS-CoV-2 S2 subunit protein was detected using rabbit anti-spike/S2 polyclonal antibody (Catalog No. 40590-T62, Sino Biological Inc.) and mouse anti-rabbit IgG peroxidase conjugate.

    Techniques: Cell Culture, Incubation, Western Blot, Reverse Transcription Polymerase Chain Reaction, Marker

    Furin cleavage score analysis of CoV S1/S2 cleavage sites. CoV S sequences were analyzed using the ProP 1 1.0 and PiTou 2 3.0 furin prediction algorithm, generating a score with green numbers indicating predicted furin cleavage and red numbers indicating no predicted furin cleavage. Purple lines denote the position of the predicted S1/S2 cleavage site. Basic arginine (R) and lysine (K) residues are highlighted in blue. Sequence numbers refer to position of amino acids within the spike protein. *For Bat-RmYN02, sequence number was determined from S alignment with SARS-CoV-2 S using Geneious. Sequences corresponding to the S1/S2 region of HCoV-HKU1 (AAT98580.1), SARS-CoV (AAT74874.1), SARS-CoV-2 (QHD43416.1), Bat-CoVRaTG13 (QHR63300.2), Bat-SL-CoVZC45 (AVP78031.1), Bat-SL-CoVZXC21 (AVP78042.1), MERS-CoV (AFS88936.1), BatCoV-HKU4 (YP_001039953.1), BatCoV-HKU5 (YP_001039962.1), and Influenza A/Chicken/Hong Kong/822.1/01/H5N1 (AF509026.2) were obtained from GenBank. Sequences corresponding to the S1/S2 and S2’ region of RmYN02 (EPI_ISL_412977) were obtained from GISAID.

    Journal: bioRxiv

    Article Title: Proteolytic activation of the SARS-CoV-2 spike S1/S2 site: a re-evaluation of furin cleavage

    doi: 10.1101/2020.10.04.325522

    Figure Lengend Snippet: Furin cleavage score analysis of CoV S1/S2 cleavage sites. CoV S sequences were analyzed using the ProP 1 1.0 and PiTou 2 3.0 furin prediction algorithm, generating a score with green numbers indicating predicted furin cleavage and red numbers indicating no predicted furin cleavage. Purple lines denote the position of the predicted S1/S2 cleavage site. Basic arginine (R) and lysine (K) residues are highlighted in blue. Sequence numbers refer to position of amino acids within the spike protein. *For Bat-RmYN02, sequence number was determined from S alignment with SARS-CoV-2 S using Geneious. Sequences corresponding to the S1/S2 region of HCoV-HKU1 (AAT98580.1), SARS-CoV (AAT74874.1), SARS-CoV-2 (QHD43416.1), Bat-CoVRaTG13 (QHR63300.2), Bat-SL-CoVZC45 (AVP78031.1), Bat-SL-CoVZXC21 (AVP78042.1), MERS-CoV (AFS88936.1), BatCoV-HKU4 (YP_001039953.1), BatCoV-HKU5 (YP_001039962.1), and Influenza A/Chicken/Hong Kong/822.1/01/H5N1 (AF509026.2) were obtained from GenBank. Sequences corresponding to the S1/S2 and S2’ region of RmYN02 (EPI_ISL_412977) were obtained from GISAID.

    Article Snippet: SARS-CoV-2 and SARS-CoV S was detected using a rabbit polyclonal antibody that recognizes the S2 domain (Cat: 40590-T62, Sinobiological) and an AlexaFluor 488 anti-rabbit secondary antibody.

    Techniques: Sequencing

    Predicted structural model of the SARS-CoV and SARS-CoV-2 S proteins. ( Inset ) Magnification of S1/S2 site with conserved R and S residues (red ribbon) and the unique four amino acid insertion P-R-R-A for SARS-CoV-2 (blue ribbon) are shown. The P’s denote the position of that amino acid from the S1/S2 cleavage site, with P1-P5 referring to amino acids before the cleavage site and P1’ referring to amino acids after the cleavage site.

    Journal: bioRxiv

    Article Title: Proteolytic activation of the SARS-CoV-2 spike S1/S2 site: a re-evaluation of furin cleavage

    doi: 10.1101/2020.10.04.325522

    Figure Lengend Snippet: Predicted structural model of the SARS-CoV and SARS-CoV-2 S proteins. ( Inset ) Magnification of S1/S2 site with conserved R and S residues (red ribbon) and the unique four amino acid insertion P-R-R-A for SARS-CoV-2 (blue ribbon) are shown. The P’s denote the position of that amino acid from the S1/S2 cleavage site, with P1-P5 referring to amino acids before the cleavage site and P1’ referring to amino acids after the cleavage site.

    Article Snippet: SARS-CoV-2 and SARS-CoV S was detected using a rabbit polyclonal antibody that recognizes the S2 domain (Cat: 40590-T62, Sinobiological) and an AlexaFluor 488 anti-rabbit secondary antibody.

    Techniques:

    Characterization of MLVpp system. ( A and B ) MLVpp infectivity in Vero E6 and Calu-3 cells. Cells were infected with MLVpps exhibiting the SARS-CoV-2 S, SARS-CoV S, VSV G or no envelope protein and assessed for luciferase activity. Error bars represent the standard error measurements of three biological replicates (n=3). ( C ) Western blot analysis of SARS-2pp and SARSpp S content. S was detected using a rabbit antibody against the SARS-CoV-2 S2 region that cross reacts against the SARS-CoV S. MLV content was detected using a mouse antibody against MLVp30. Image cropped from a singular Western blot and lanes are reshown in Figure 3C and 5C .

    Journal: bioRxiv

    Article Title: Proteolytic activation of the SARS-CoV-2 spike S1/S2 site: a re-evaluation of furin cleavage

    doi: 10.1101/2020.10.04.325522

    Figure Lengend Snippet: Characterization of MLVpp system. ( A and B ) MLVpp infectivity in Vero E6 and Calu-3 cells. Cells were infected with MLVpps exhibiting the SARS-CoV-2 S, SARS-CoV S, VSV G or no envelope protein and assessed for luciferase activity. Error bars represent the standard error measurements of three biological replicates (n=3). ( C ) Western blot analysis of SARS-2pp and SARSpp S content. S was detected using a rabbit antibody against the SARS-CoV-2 S2 region that cross reacts against the SARS-CoV S. MLV content was detected using a mouse antibody against MLVp30. Image cropped from a singular Western blot and lanes are reshown in Figure 3C and 5C .

    Article Snippet: SARS-CoV-2 and SARS-CoV S was detected using a rabbit polyclonal antibody that recognizes the S2 domain (Cat: 40590-T62, Sinobiological) and an AlexaFluor 488 anti-rabbit secondary antibody.

    Techniques: Infection, Luciferase, Activity Assay, Western Blot

    The levels of SARS-CoV-2-reactive to S1 and S2 subunits (S1 + S2) and nucleocapsid in human milk collected during the COVID-19 pandemic. The levels of a S1 + S2- and b nucleocapsid-reactive secretory IgM (SIgM)/IgM, IgG, and secretory IgA (SIgA)/IgA in human milk. Values are mean ± SD, n = 41 for women. Asterisks show statistically significant differences between variables (*** p

    Journal: Journal of Perinatology

    Article Title: Difference in levels of SARS-CoV-2 S1 and S2 subunits- and nucleocapsid protein-reactive SIgM/IgM, IgG and SIgA/IgA antibodies in human milk

    doi: 10.1038/s41372-020-00805-w

    Figure Lengend Snippet: The levels of SARS-CoV-2-reactive to S1 and S2 subunits (S1 + S2) and nucleocapsid in human milk collected during the COVID-19 pandemic. The levels of a S1 + S2- and b nucleocapsid-reactive secretory IgM (SIgM)/IgM, IgG, and secretory IgA (SIgA)/IgA in human milk. Values are mean ± SD, n = 41 for women. Asterisks show statistically significant differences between variables (*** p

    Article Snippet: SARS-CoV-2 S1 + S2- and nucleocapsid-reactive antibodies The levels of SARS-CoV-2 S1 + S2 and nucleocapsid-reactive SIgM/IgM, IgG and SIgA/IgA were determined using ELISAs that were adapted from our previous publications [ , – ].

    Techniques:

    Regression linear between antibodies reactive to SARS-CoV-2 S1 and S2 subunits (S1 + S2) and nucleocapsid in human milk collected during the COVID-19 pandemic in 41 women. a Positive correlation of S1 + S2-reactive secretory IgA (SIgA)/IgA and S1 + S2-reactive secretory IgM (SIgM)/IgM. b Positive correlation between S1 + S2-reactive SIgA/IgA and nucleocapsid-reactive SIgA/IgA. c Positive correlation between nucleocapsid-reactive SIgA/IgA and SIgM/IgM. d Positive correlation between nucleocapsid-reactive SIgM/IgM and S1 + S2-reactive SIgM/IgM. Pearson correlation coefficients ( r ) were determined when p

    Journal: Journal of Perinatology

    Article Title: Difference in levels of SARS-CoV-2 S1 and S2 subunits- and nucleocapsid protein-reactive SIgM/IgM, IgG and SIgA/IgA antibodies in human milk

    doi: 10.1038/s41372-020-00805-w

    Figure Lengend Snippet: Regression linear between antibodies reactive to SARS-CoV-2 S1 and S2 subunits (S1 + S2) and nucleocapsid in human milk collected during the COVID-19 pandemic in 41 women. a Positive correlation of S1 + S2-reactive secretory IgA (SIgA)/IgA and S1 + S2-reactive secretory IgM (SIgM)/IgM. b Positive correlation between S1 + S2-reactive SIgA/IgA and nucleocapsid-reactive SIgA/IgA. c Positive correlation between nucleocapsid-reactive SIgA/IgA and SIgM/IgM. d Positive correlation between nucleocapsid-reactive SIgM/IgM and S1 + S2-reactive SIgM/IgM. Pearson correlation coefficients ( r ) were determined when p

    Article Snippet: SARS-CoV-2 S1 + S2- and nucleocapsid-reactive antibodies The levels of SARS-CoV-2 S1 + S2 and nucleocapsid-reactive SIgM/IgM, IgG and SIgA/IgA were determined using ELISAs that were adapted from our previous publications [ , – ].

    Techniques:

    The levels of antibodies reactive to SARS-CoV-2 S1 and S2 subunits (S1 + S2) and nucleocapsid in human milk collected during the COVID-19 pandemic (2020-HM) and 2 years prior this pandemic (2018-HM). Levels of S1 + S2-reactive a secretory IgM (SIgM)/IgM, b IgG and c secretory IgA (SIgA)/IgA in 2020-HM and 2018-HM. Levels of nucleocapsid-reactive d SIgM/IgM, e IgG and f SIgA/IgA in 2020-HM and 2018-HM. Values are mean ± SD, n = 41 for 2020-HM and n = 16 for 2018-HM. Asterisks show statistically significant differences between variables (* p

    Journal: Journal of Perinatology

    Article Title: Difference in levels of SARS-CoV-2 S1 and S2 subunits- and nucleocapsid protein-reactive SIgM/IgM, IgG and SIgA/IgA antibodies in human milk

    doi: 10.1038/s41372-020-00805-w

    Figure Lengend Snippet: The levels of antibodies reactive to SARS-CoV-2 S1 and S2 subunits (S1 + S2) and nucleocapsid in human milk collected during the COVID-19 pandemic (2020-HM) and 2 years prior this pandemic (2018-HM). Levels of S1 + S2-reactive a secretory IgM (SIgM)/IgM, b IgG and c secretory IgA (SIgA)/IgA in 2020-HM and 2018-HM. Levels of nucleocapsid-reactive d SIgM/IgM, e IgG and f SIgA/IgA in 2020-HM and 2018-HM. Values are mean ± SD, n = 41 for 2020-HM and n = 16 for 2018-HM. Asterisks show statistically significant differences between variables (* p

    Article Snippet: SARS-CoV-2 S1 + S2- and nucleocapsid-reactive antibodies The levels of SARS-CoV-2 S1 + S2 and nucleocapsid-reactive SIgM/IgM, IgG and SIgA/IgA were determined using ELISAs that were adapted from our previous publications [ , – ].

    Techniques:

    Percentage of detected antibodies reactive to S1 and S2 subunits (S1 + S2), and nucleocapsid from SARS-CoV-2 in human milk collected during the COVID-19 pandemic. S1+S2-reactive a secretory IgM (SIgM)/IgM, b IgG and c secretory IgA (SIgA)/IgA in human milk from 41 women. Nucleocapsid-reactive d SIgM/IgM, e IgG and f SIgA/IgA. Milk expression period was from 30/02/20 to 03/04/20 in the United States.

    Journal: Journal of Perinatology

    Article Title: Difference in levels of SARS-CoV-2 S1 and S2 subunits- and nucleocapsid protein-reactive SIgM/IgM, IgG and SIgA/IgA antibodies in human milk

    doi: 10.1038/s41372-020-00805-w

    Figure Lengend Snippet: Percentage of detected antibodies reactive to S1 and S2 subunits (S1 + S2), and nucleocapsid from SARS-CoV-2 in human milk collected during the COVID-19 pandemic. S1+S2-reactive a secretory IgM (SIgM)/IgM, b IgG and c secretory IgA (SIgA)/IgA in human milk from 41 women. Nucleocapsid-reactive d SIgM/IgM, e IgG and f SIgA/IgA. Milk expression period was from 30/02/20 to 03/04/20 in the United States.

    Article Snippet: SARS-CoV-2 S1 + S2- and nucleocapsid-reactive antibodies The levels of SARS-CoV-2 S1 + S2 and nucleocapsid-reactive SIgM/IgM, IgG and SIgA/IgA were determined using ELISAs that were adapted from our previous publications [ , – ].

    Techniques: Expressing