Anti-ACE2 antibody [RM1070]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACE2 antibody [RM1070] (AB314771)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling ACE2 with ab314771 at 1/5000 (0.109 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on human liver. The section was incubated with ab314771 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 was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACE2 antibody [RM1070] (AB314771)

Immunohistochemical analysis of paraffin-embedded human testis tissue labeling ACE2 with ab314771 at 1/5000 (0.109 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human testis. The section was incubated with ab314771 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 was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACE2 antibody [RM1070] (AB314771)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling ACE2 with ab314771 at 1/5000 (0.109 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human kidney. The section was incubated with ab314771 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 was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-ACE2 antibody [RM1070] (AB314771)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HEK-293T (human epithelial cell line from embryonic kidney) transfected with myc-tagged ACE2 expression vector cells labelling ACE2 with ab314771 at 1/500 (1.092 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing cytoplasmic staining in 293T cells transfected with ACE2 protein expression vector containing a myc tag, no staining on 293T transfected with empty vector containing a myc-tag. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab223894 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/100 5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-ACE2 antibody [RM1070] (AB314771)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized Calu-3 (human lung adenocarcinoma epithelial cell) cells labelling ACE2 with ab314771 at 1/50 (10.92 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing cytoplasmic staining in Calu-3 cell line. Negative control : HeLa.Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.

• IP

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Immunoprecipitation - Anti-ACE2 antibody [RM1070] (AB314771)

ACE2 was immunoprecipitated from 0.35 mg Calu-3 (human lung adenocarcinoma epithelial cell) whole cell lysate with ab314771 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314771 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Calu-3 (human lung adenocarcinoma epithelial cell) whole cell lysate Lane 2 : ab314771 IP in Calu-3 (human lung adenocarcinoma epithelial cell) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab314771 in Calu-3 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-ACE2 antibody [RM1070] (ab314771) at 1/30 dilution

All lanes:

Calu-3 (human lung adenocarcinoma epithelial cell) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 41s

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACE2 antibody [RM1070] (AB314771)

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling ACE2 with ab314771 at 1/2000 (0.273 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse kidney. The section was incubated with ab314771 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 was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

• WB

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Western blot - Anti-ACE2 antibody [RM1070] (AB314771)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Negative control : liver (PMID : 10924499) and HeLa (PMID : 32926098). In western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution. Exposure time : Lanes 1-4 : 180 seconds; Lane 5 : 15 seconds

All lanes:

Western blot - Anti-ACE2 antibody [RM1070] (ab314771) at 1/1000 dilution

Lane 1:

Human testis tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Calu-3 (human lung adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

Human kidney tissue lysate at 20 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Observed band size: 110 kDa

false

• WB

Supplier Data

Western blot - Anti-ACE2 antibody [RM1070] (AB314771)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Negative control : liver (PMID : 10924499). In western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-ACE2 antibody [RM1070] (ab314771) at 1/1000 dilution

Lane 1:

Mouse kidney tissue lysate at 20 µg

Lane 2:

Mouse liver tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 110 kDa

false

Exposure time: 180s

• I-ELISA

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Indirect ELISA - Anti-ACE2 antibody [RM1070] (AB314771)

Indirect ELISA analysis of ab314771 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-RABbit IgG (H+L) at 1 : 2500 dilution. Antigen : Human ACE2. Antigen concentration : 1000 ng/ml

• WB

Supplier Data

Western blot - Anti-ACE2 antibody [RM1070] (AB314771)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. In western blot, anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution. In western blot, anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-ACE2 antibody [RM1070] (ab314771) at 1/1000 dilution

Lane 1:

293T cells transfected with an empty vector containing a myc-his tag, whole cell lysate at 4 µg

Lane 2:

293T cells transfected with a human ACE2 expression vector containing a myc-his-tag, whole cell lysate at 4 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 110 kDa

false

Exposure time: 4s