recombinant mouse ace2  (R&D Systems)

Journal: Frontiers in Pharmacology

Article Title: SGLT2i and GLP1-RA exert additive cardiorenal protection with a RAS blocker in uninephrectomized db/db mice

doi: 10.3389/fphar.2024.1415879

Figure Lengend Snippet: ACE2/ACE ratios in the kidney cortex and serum of vehicle-treated db/m mice, vehicle-treated db/db mice, and db/db mice treated with empagliflozin, semaglutide, or their combination with ramipril. (A) ACE2 and ACE mRNA expression and (B) ACE2/ACE activity ratios in the kidney cortex. (C) ACE2/ACE activity ratio in serum. Individual values of ACE and ACE2 gene expression and activity are shown in . db/m: non-diabetic mice treated with vehicle. db/db: diabetic mice treated with vehicle. db/db EMP + RAM: diabetic mice treated with empagliflozin and ramipril. db/db SEM + RAM: diabetic mice treated with semaglutide and ramipril. db/db EMP + SEM + RAM: diabetic mice treated with empagliflozin, semaglutide, and ramipril. $ p < 0.05 vehicle-treated db/db mice compared to vehicle-treated non-diabetic mice. * p < 0.05 db/db mice treated with empagliflozin, semaglutide, or their combination compared to vehicle-treated db/db mice.

Article Snippet: As positive and negative controls, 0.008 ng/μL of recombinant mouse ACE2 (3437-ZN-010, R&D Systems) and lysis buffer alone were used, respectively.

Techniques: Expressing, Activity Assay

Journal: bioRxiv

Article Title: SARS-CoV-2 infects neurons, astrocytes, choroid plexus epithelial cells and pericytes of the human central nervous system

doi: 10.1101/2023.11.21.568132

Figure Lengend Snippet: Primary human astrocytes, neurons and choroid plexus epithelial cells were incubated with ACE2 neutralizing antibody for 1h (10 μg/mL) before being inoculated with SARS-CoV-2 (Italy_INMI1) at an MOI of 0.1 for 1h. A) 72h post-infection, cells were fixed and immunolabelled for anti-SARS-CoV-2 spike protein, and infected cells enumerated. Data are presented as % neutralization relative to the untreated control. B) Cells were lysed and SARS-CoV-2 nucleocapsid (N1) gene quantified by qRT-PCR. Data are presented as SARS-CoV-2 N1 genome copies/mL (gc/mL). ****p<0.0001, **p<0.01, *p<0.05, ns=not significant. N=3 independent experiments.

Article Snippet: Primary antibodies used for immunofluorescent staining and neutralisation assays were as follows: SARS-CoV-2 Spike mAb anti-rabbit IgG (SinoBiological, Beijing, China); dsRNA K1 mAb anti-mouse IgG2a (Scicons, Susteren, The Netherlands), and recombinant anti-ACE2 neutralising mAb anti-mouse IgG1 (SinoBiological, Beijing, China).

Techniques: Incubation, Infection, Neutralization, Quantitative RT-PCR

Journal: NPJ Vaccines

Article Title: Effective vaccination strategy using SARS-CoV-2 spike cocktail against Omicron and other variants of concern

doi: 10.1038/s41541-022-00580-z

Figure Lengend Snippet: Schematic map of SARS-CoV-2 S protein of the original wild-type (WT-S) strain ( a ) and Omicron BA1 subvariant (BA1-S) ( b ). Mutant amino acid residues of Omicron BA1 subvariant are shown in the S1 (including N-terminal domain (NTD) and receptor-binding domain (RBD)) and S2 subunits of S protein, respectively. Mutant amino acid residues in the RBD of Omicron BA2, BA2.12.1, and BA5 are shown ( b ). SP signal peptide. FP fusion peptide. HR1 and HR2, heptad repeat regions 1 and 2. TM transmembrane domain. CP cytoplasmic tail. ELISA analysis of binding of Omicron BA1-S protein (BA1-S) or original S protein (WT-S) to human angiotensin-converting enzyme 2 (hACE2) ( c ), hamster ACE2 ( d ), and mouse ACE2 ( e ) proteins, respectively. Control, PBS. Statistical significance between the binding of WT-S and BA1-S proteins to mouse ACE2 protein was analyzed using a two-tailed student t -test, and * ( P < 0.05) indicates a significant difference. f Flow cytometry analysis of binding of BA1-S (blue line) and WT-S (red line) proteins to bat ACE2-expressing 293T cells (bat-ACE2/293T). 293T cells were transiently transfected with bat ACE2 plasmid and incubated with each protein (5 µg/ml) for analysis by flow cytometry. Gray shading, mock-incubated cells. MFI, median fluorescence intensity. ELISA for detection of binding of WT-S and BA1-S proteins to SARS-CoV-2 S-vaccinated human ( g ) and mouse ( h ) serum-neutralizing antibodies, respectively. The data (in c – h ) are expressed as mean ± standard deviation of the mean (s.e.m.) of the duplicate to quadruple wells. The experiments were repeated twice, leading to similar results.

Article Snippet: For binding to mouse ACE2 and hamster ACE2, ELISA plates were coated with respective mouse ACE2 (1 μg/ml, R&D System 3437-ZN) or hamster ACE2 (1 μg/ml, R&D System 10578-ZN) protein, and then sequentially incubated with serial dilutions of each S protein, SARS-CoV-2 S-specific mouse polyclonal antibody (1:2,000 dilution, Laboratory stock), and anti-mouse IgG-Fab-HRP antibody (1:5000 dilution, Sigma A9917-1ML) for 1 h at 37 °C.

Techniques: Mutagenesis, Binding Assay, Enzyme-linked Immunosorbent Assay, Control, Two Tailed Test, Flow Cytometry, Expressing, Transfection, Plasmid Preparation, Incubation, Fluorescence, Standard Deviation