Ace2 af933

Manufactured by R&D Systems

ACE2 (AF933) is a recombinant human Angiotensin-Converting Enzyme 2 (ACE2) protein. ACE2 is a type I integral membrane protein that functions as a receptor for the SARS-CoV-2 virus. The ACE2 protein provided in this product is a truncated version containing only the extracellular domain.

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Lab products found in correlation

Gapdh, by GenScript (1 mentions) Cd83 hb15e, by BD (1 mentions) Cd86 2331 fun 1, by BD (1 mentions) Donkey anti goat a21447, by Thermo Fisher Scientific (1 mentions) Bsa pbs, by Merck Group (1 mentions) Cd80 l307.4, by BD (1 mentions) Facscanto flow cytometer, by BD (1 mentions)

2 protocols using ace2 af933

Protein Expression Analysis by Western Blot

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Protein expression was analyzed by polyacrylamide gel electrophoresis on 8% to 16% precast gradient gels (Thermo) and Western blotting using antibodies to ACE2 (AF933; R&D Systems), the c-Myc epitope (clone 9E10; Santa Cruz Biotechnology), SARS spike (NB100-56578; Novus Biologicals), HIV-1 Gag p24 (clone 749140; R&D), and GAPDH (glyceraldehyde-3-phosphate dehydrogenase; GenScript) in NETT-G (150 mM NaCl, 5 mM EDTA, 50 mM Tris, 0.05% Triton X-100, 0.25% gelatin, pH 7.5) and donkey anti-mouse horseradish peroxidase (HRP)-coupled (Dianova), goat anti-rabbit HRP-coupled (Life Technologies), or rabbit anti-goat HRP-coupled (Proteintech) secondary antibody in 5% dry milk powder in PBS with 0.05% Tween 20. Imaging was performed using the Immobilon Forte substrate (Merck) on an INTAS ECL ChemoCam system.

Hörnich B.F., Großkopf A.K., Schlagowski S., Tenbusch M., Kleine-Weber H., Neipel F., Stahl-Hennig C, & Hahn A.S. (2021). SARS-CoV-2 and SARS-CoV Spike-Mediated Cell-Cell Fusion Differ in Their Requirements for Receptor Expression and Proteolytic Activation. Journal of Virology, 95(9), e00002-21.

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Cell Surface Marker Analysis by Flow Cytometry

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For cell surface staining, cells were incubated in 0.5% PBS‐BSA (Sigma‐Aldrich) containing antibodies for 30 min at 4°C. Single‐cell measurements were performed on a FACS Canto flow cytometer (BD Biosciences) and FlowJo V10 software (TreeStar) was used to analyze the data. The antibody clones used are: CD86 (2331 (FUN‐1), BD Pharmingen), CD80 (L307.4, BD Pharmingen), CD83 (HB15e, BD Pharmingen), ACE2 (AF933, R&D Systems), goat‐IgG (AB‐2535864, ThermoFisher Scientific), donkey‐anti‐goat (A‐21447, ThermoFisher Scientific). For each experiment, live cells were gated on FSC and SSC and analyzed further with the markers mentioned (Supporting information Fig. S1). The authors adhered to the guidelines for the use of flow cytometry and cell sorting in immunological studies [37 (link)].

van der Donk L.E., Eder J., van Hamme J.L., Brouwer P.J., Brinkkemper M., van Nuenen A.C., van Gils M.J., Sanders R.W., Kootstra N.A., Bermejo‐Jambrina M, & Geijtenbeek T.B. (2022). SARS‐CoV‐2 infection activates dendritic cells via cytosolic receptors rather than extracellular TLRs. European Journal of Immunology, 52(4), 646-655.

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