goat anti-hace2 polyclonal antibody  (R&D Systems)

Journal: JCI insight

Article Title: Highly susceptible SARS-CoV-2 model in CAG promoter-driven hACE2-transgenic mice.

doi: 10.1172/jci.insight.152529

Figure Lengend Snippet: Figure 1. The pathogenesis of SARS-CoV-2 infection is exacerbated in a viral dose-dependent manner. (A) Western blot of human ACE2 protein using various organs in WT and CAG-hACE2 mice. The top and bottom rows show hACE2 and GAPDH, respectively. (B) Schematic diagram of experimental schedule. Male and female C57BL/6 and CAG-hACE2 mice were infected via respiratory tract with SARS-CoV-2 (2 × 102 TCID50, n = 5; 2 × 103 TCID50, n = 6; 2 × 104 TCID50, n = 5) and were administrated with an equal volume of PBS for mock infection controls (WT, n = 3; CAG-hACE2 mice, n = 3). Body weight and survival were recorded daily for up to 14 days. (C and D) Percentage of initial body weight (C) and survival rate (D). Numbers in C represent the number of mice measured for body weight at each time. White and black circles indicate WT with mock infection and WT infected with 2 × 104 TCID50, respectively. Triangles denote CAG-hACE2 mice. White, orange, blue, and red triangles represent PBS, 2 × 102 TCID50, 2 × 103 TCID50, and 2 × 104 TCID50, respectively.

Article Snippet: Then, the membranes were incubated with HRP-conjugated goat anti-mouse polyclonal IgG for hACE2 (1:2000, catalog 172-1011, Bio-Rad) or HRP-conjugated goat anti-rabbit polyclonal IgG for GAPDH (1:2000, catalog 172-1019, Bio-Rad) for 1 hour at room temperature, and the protein signals were detected using a chemiluminescence detection reagent (Chemi-Lumi One L; Nacalai Tesque Inc.) and visualized using the ImageQuant LAS 4000 system (Cytiva).

Techniques: Infection, Western Blot

Journal: JCI insight

Article Title: Highly susceptible SARS-CoV-2 model in CAG promoter-driven hACE2-transgenic mice.

doi: 10.1172/jci.insight.152529

Figure Lengend Snippet: Figure 7. Characterization of CAG-promoter hACE2-transgenic mice. (A) Chromosome map of annotated hACE2-containing reads. The pink horizontal bars indicate the number of annotated ACE2-containing reads. Annotated site of integrated hACE2 is indicated as a blue box. (B) Detail of annotated site in chromosome 1. The horizontal red and pink bars indicate the ACE2-containing reads mapped to the plus and minus strand of the mouse reference genome (GRCh38/mm10), respectively. The blue box represents the duplicated region. (C) Schematic diagrams and sequences are shown as the hACE2- integrated site inside the first intron region of the Colgalt2 gene locus. The blue boxes denote duplicated regions. Each yellow arrow indicates the entire hACE2 sequence. Consensus sequences of reads at 5′- and 3′-junction sites are shown.

Article Snippet: Then, the membranes were incubated with HRP-conjugated goat anti-mouse polyclonal IgG for hACE2 (1:2000, catalog 172-1011, Bio-Rad) or HRP-conjugated goat anti-rabbit polyclonal IgG for GAPDH (1:2000, catalog 172-1019, Bio-Rad) for 1 hour at room temperature, and the protein signals were detected using a chemiluminescence detection reagent (Chemi-Lumi One L; Nacalai Tesque Inc.) and visualized using the ImageQuant LAS 4000 system (Cytiva).

Techniques: Transgenic Assay, Sequencing

Journal: JCI insight

Article Title: Highly susceptible SARS-CoV-2 model in CAG promoter-driven hACE2-transgenic mice.

doi: 10.1172/jci.insight.152529

Figure Lengend Snippet: Figure 8. Protection against SARS-CoV-2 by immunization of SARS-CoV-2 RBD protein. (A) Schematic showing the experimental schedule. Male and female CAG-hACE2 mice were immunized with RBD-mFc/AddaVax twice before infection and were infected via respiratory tract with SARS-CoV-2 (2 × 103 TCID50, n = 4; 1 × 104 TCID50, n = 4). PBS/AddaVax–immunized CAG-hACE2 mice were used as a control (2 × 103 TCID50, n = 6; 1 × 104 TCID50, n = 6). (B) Expression of RBD-sIgG in plasma from PBS/AddaVax or RBD-mFc/AddaVax–immunized CAG-hACE2 mice at 2 days before infection. Blue and red indicate 2 × 103 TCID50 and 1 × 104 TCID50, respectively. (C and D) Percentage of initial body weight (C) and survival rate (D). Numbers in C represent the number of mice measured for body weight at each time. White and colored circles indicate mice injected with PBS/AddaVax or RBD-mFc/AddaVax, respectively. Blue and red denote infection doses of 2 × 103 TCID50 and 1 × 104 TCID50, respectively. Data are presented as the mean (B) and the mean ± SEM (C). Statistical analyses were performed using 2-tailed unpaired t test for body weight loss (B and C) and log-rank (mantel-cox) test for survival rate (D). P < 0.05 for comparison with RBD-mFc/AddaVax (2 × 103 TCID50) or PBS/AddaVax–immunized mice, respectively.

Article Snippet: Then, the membranes were incubated with HRP-conjugated goat anti-mouse polyclonal IgG for hACE2 (1:2000, catalog 172-1011, Bio-Rad) or HRP-conjugated goat anti-rabbit polyclonal IgG for GAPDH (1:2000, catalog 172-1019, Bio-Rad) for 1 hour at room temperature, and the protein signals were detected using a chemiluminescence detection reagent (Chemi-Lumi One L; Nacalai Tesque Inc.) and visualized using the ImageQuant LAS 4000 system (Cytiva).

Techniques: Infection, Control, Expressing, Clinical Proteomics, Injection, Comparison

Journal: JCI insight

Article Title: Highly susceptible SARS-CoV-2 model in CAG promoter-driven hACE2-transgenic mice.

doi: 10.1172/jci.insight.152529

Figure Lengend Snippet: Figure 9. Protection against SARS-CoV-2 infection by the injection of RBD-Fc–immunized mouse plasma. (A) Schematic showing the experimental schedule. Male and female CAG-hACE2 mice were intravenously injected with pooled plasma from RBD-mFc/AddaVax–immunized mice at doses of 15 μg/shot (n = 6), 50 μg/shot (n = 5), and 150 μg/shot (n = 6) 1 day before SARS-CoV-2 infection and were infected via respiratory tract with SARS-CoV-2 at a dose of 1 × 104 TCID50. CAG-hACE2 mice that were injected with PBS-treated pooled plasma were used as a control (n = 6). (B) The measurement of SARS-CoV-2–neutralizing antibodies in pooled plasma. Black circles and white, violet, and gray diamonds indicate the standard, pooled plasma from PBS/ AddaVax–treated mice, pooled plasma from RBD-mFc/AddaVax–immunized mice, and mouse neutralizing antibody, respectively. (C and D) Percentage of initial body weight (C) and survival rate (D). Numbers in C represent the number of mice measured for body weight at each time. Black circles and orange, blue, and red triangles indicate plasma from PBS/AddaVax–immunized mice and RBD-mFc/AddaVax–immunized mice at doses of 150 ng/mL (n = 6), 500 ng/mL (n = 5), and 1500 ng/mL (n = 6), respectively. Data were presented as the mean (B) and the mean ± SEM (C). Statistical analyses were performed using log-rank (mantel-cox) test for survival rate (D). P < 0.05 for the comparison with PBS-treated pool plasma.

Article Snippet: Then, the membranes were incubated with HRP-conjugated goat anti-mouse polyclonal IgG for hACE2 (1:2000, catalog 172-1011, Bio-Rad) or HRP-conjugated goat anti-rabbit polyclonal IgG for GAPDH (1:2000, catalog 172-1019, Bio-Rad) for 1 hour at room temperature, and the protein signals were detected using a chemiluminescence detection reagent (Chemi-Lumi One L; Nacalai Tesque Inc.) and visualized using the ImageQuant LAS 4000 system (Cytiva).

Techniques: Infection, Injection, Clinical Proteomics, Control, Comparison