Anti ace2 recombinant rabbit monoclonal antibody

HEK293T cells were seeded in a μ-Slide 8-Well Chamber Slide (iBidi GmbH) after precoating with poly-l-lysine (Cultrex). After 24 h, the cells were transfected with the pCAGGS encoding FLAG-tagged ACE2 proteins or empty pCAGGS. At 24 h posttransfection, the cells were washed with PBS and fixed with PBS containing 4% paraformaldehyde for 15 min. After a wash with PBS, the cells were incubated with PBS containing 3% bovine serum albumin for blocking for 1 h at room temperature. The cells were washed three times with PBST and then incubated with an anti-ACE2 recombinant rabbit monoclonal antibody (Invitrogen, SN0754) recognizing an epitope at amino acid positions 190 to 230 as the primary antibody for 1 h at room temperature. The cells were washed with PBST and then incubated with an Alexa Fluor 488-conjugated goat anti-rabbit IgG antibody (Molecular Probes) as a secondary antibody and counterstained with 1 μg/mL 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI; Molecular Probes) for 1 h in the dark at room temperature. Images were acquired with a 63× oil lens objective on a Zeiss LSM700 inverted microscope using ZEN 2009 software (Carl Zeiss).

HEK293T cells were seeded in 6-well plates (2.0 × 10⁴ cells per well) precoated with poly-l-lysine (Cultrex). After 24 h, the cells were transfected with pCAGGS encoding FLAG-tagged ACE2 WT or SNV mutant proteins using TransIT-LT1. At 24 h posttransfection, these cells were washed with PBS and detached using 0.25% trypsin. Cells were fixed with PBS containing 4% paraformaldehyde for 15 min. After a wash with PBS, cells were incubated with an anti-ACE2 recombinant rabbit monoclonal antibody (Invitrogen, SN0754) for 1 h at room temperature. The cells were next stained with the Alexa Fluor 488-conjugated goat anti-rabbit IgG antibody (Molecular Probes) for 30 min at 4°C in the dark. After two washes, the surface expression of the exogenous ACE2 proteins was analyzed by using a FACSCanto flow cytometer (BD Biosciences) and FlowJo software (Tree Star).

HEK293T cells were seeded in 6-well plates (2.0 × 10⁴ cells per well) precoated with poly-l-lysine (Cultrex). After 24 h, the cells were transfected with pCAGGS encoding FLAG-tagged ACE2 WT or SNV mutant proteins using TransIT-LT1. At 24 h posttransfection, these cells were washed with PBS and detached using 0.25% trypsin. Cells were fixed with PBS containing 4% paraformaldehyde for 15 min. After a wash with PBS, cells were incubated with an anti-ACE2 recombinant rabbit monoclonal antibody (Invitrogen, SN0754) for 1 h at room temperature. The cells were next stained with the Alexa Fluor 488-conjugated goat anti-rabbit IgG antibody (Molecular Probes) for 30 min at 4°C in the dark. After two washes, the surface expression of the exogenous ACE2 proteins was analyzed by using a FACSCanto flow cytometer (BD Biosciences) and FlowJo software (Tree Star).