Recombinant Anti-ACE2 antibody [EPR24705-45] (ab272500)

ab272500 staining ACE2 in wild-type Caco2 cells (top panel) and ACE2 knockout Caco2 cells (ab273731) (bottom panel). The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab272500 at 10μg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor^® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor^® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

Immunohistochemical analysis of paraffin-embedded Human testis tissue labelling ACE2 with ab272500 at 1/100 (5.0 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection). Membranous and cytoplasmic staining in human testis. The section was incubated with ab272500 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond^® Polymer Refine Detection) .

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T cells labelling ACE2 with ab272500 at 1/250 (2.0 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing positive staining in HEK-293T cells transfected with myc-tagged ACE2 expression vector. Myc-Tag Mouse mAb (Alexa Fluor^® 647) was used at 1/100 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Calu-3 cells labelling ACE2 with ab272500 at 1/50 (10.0 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in subsets of Calu-3 cells. Negative control: HeLa (PMID: 32926098). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) (Red) was used at 1/200 dilution. The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) at 1/1000 dilution.

Flow cytometric analysis of HeLa (Human cervix adenocarcinoma epithelial cell, Left)/ Calu-3 (human lung adenocarcinoma epithelial cell, Right) cells labelling ACE2 with ab272500 at 1/500 dilution (0.1ug). Goat anti rabbit IgG (Alexa Fluor^® 647, ab150079) at 1/2000 dilution was used as the secondary antibody.

Negative control: HeLa (PMID: 32926098). Gated on viable cells.

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

The expression profile/ molecular weight  observed is consistent with what has been described in the literature (PMID:33008593).

Negative control: HeLa (PMID:32926098).

Exposure time: 3 minutes

ACE2 was immunoprecipitated from 0.35 mg Human testis lysate with ab272500 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab272500 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: Human testis lysate 10 ug

Lane 2: ab272500 IP in Human testis lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab272500 in human testis whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 32 seconds

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labelling ACE2 with ab272500 at 1/100 (5.0 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Membranous and cytoplasmic staining in human kidney. The section was incubated with ab272500 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND™ RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) .

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

The expression profile/ molecular weight  observed is consistent with what has been described in the literature (PMID:10924499, PMID: 32661139).

Negative control: liver (PMID:10924499).

Exposure time: 3 minutes

Immunohistochemical analysis of paraffin-embedded Human liver tissue labelling ACE2 with ab272500 at 1/100 (5.0 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control: No staining in human liver. The section was incubated with ab272500 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond^®Polymer Refine Detection) .

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.