Recombinant Anti-ACE2 antibody [EPR4435(2)] (ab108252)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4435(2)] to ACE2
- Suitable for: WB, IP, IHC-P, Indirect ELISA
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-ACE2 antibody [EPR4435(2)]
See all ACE2 primary antibodies -
Description
Rabbit monoclonal [EPR4435(2)] to ACE2 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IP, IHC-P, Indirect ELISAmore details
Unsuitable for: Flow Cyt or ICC/IF -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available asab198988) -
Positive control
- WB: Human testis, kidney and lung tissue lysates; Human fetal kidney tissue lysate; Calu-3, HepG2 and Caco-2 cell lysates. Human and rat heart tissue lysate; Human lung tissue lysate; Mouse and rat spleen, testis lung tissue lysate; IHC-P: Human, mouse, and rat kidney tissues. IP: Human testis tissue lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR4435(2) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab108252 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB | (5) |
1/1000 - 1/10000. Predicted molecular weight: 92 kDa.Can be blocked with ACE2 peptide (ab198988).
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IP |
1/10 - 1/100.
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IHC-P | (1) |
1/6400 - 1/32000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
For unpurified use at 1/100 - 1/250. |
Indirect ELISA |
Use at an assay dependent concentration.
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Notes |
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WB
1/1000 - 1/10000. Predicted molecular weight: 92 kDa.Can be blocked with ACE2 peptide (ab198988). |
IP
1/10 - 1/100. |
IHC-P
1/6400 - 1/32000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. For unpurified use at 1/100 - 1/250. |
Indirect ELISA
Use at an assay dependent concentration. |
Target
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Function
Carboxypeptidase which converts angiotensin I to angiotensin 1-9, a peptide of unknown function, and angiotensin II to angiotensin 1-7, a vasodilator. Also able to hydrolyze apelin-13 and dynorphin-13 with high efficiency. May be an important regulator of heart function. In case of human coronaviruses SARS and HCoV-NL63 infections, serve as functional receptor for the spike glycoprotein of both coronaviruses. -
Tissue specificity
Expressed in endothelial cells from small and large arteries, and in arterial smooth muscle cells. Expressed in lung alveolar epithelial cells, enterocytes of the small intestine, Leydig cells and Sertoli cells (at protein level). Expressed in heart, kidney, testis, and gastrointestinal system. -
Sequence similarities
Belongs to the peptidase M2 family. -
Post-translational
modificationsN-glycosylation on Asn-90 may limit SARS infectivity. -
Cellular localization
Secreted and Cell membrane. - Information by UniProt
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Database links
- Entrez Gene: 59272 Human
- Entrez Gene: 70008 Mouse
- Entrez Gene: 302668 Rat
- Omim: 300335 Human
- SwissProt: Q9BYF1 Human
- SwissProt: Q8R0I0 Mouse
- SwissProt: Q5EGZ1 Rat
- Unigene: 178098 Human
see all -
Alternative names
- ACE 2 antibody
- ACE related carboxypeptidase antibody
- ACE-related carboxypeptidase antibody
see all
Images
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All lanes : Anti-ACE2 antibody [EPR4435(2)] (ab108252) at 1/1000 dilution
Lane 1 : Human heart tissue lysate
Lane 2 : Rat heart tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 92 kDa
Observed band size: 110,120 kDa why is the actual band size different from the predicted?
Exposure time: 180 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST.
ab181602 was used as a GAPDH loading control.
Two bands observed by ab108252 corresponding to glycosylation and non-glycosylation forms.
Signal in heart tissue is low, we recommend loading more amount of lysate or using lower antibody dilution to improve result.
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All lanes : Anti-ACE2 antibody [EPR4435(2)] (ab108252) at 1/1000 dilution
Lane 1 : Human testis tissue lysate at 20 µg
Lane 2 : Human lung tissue lysate at 20 µg
Lane 3 : Mouse testis tissue lysate
Lane 4 : Mouse spleen tissue lysate
Lane 5 : Mouse lung tissue lysate
Lane 6 : Rat testis tissue lysate
Lane 7 : Rat spleen tissue lysate
Lane 8 : Rat lung tissue lysate
Secondary
All lanes : Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 92 kDa
Observed band size: 110,120 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
ab181602 was used as GAPDH loading control.
Exposure time: Lane 1: 7 seconds; Lane 2-8: 180 seconds.
Two bands observed by ab108252 corresponding to glycosylation and non-glycosylation forms.
Signal in mouse and rat tissues are low, we recommend loading more amount of lysate or using lower antibody dilution to improve result.
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All lanes : Anti-ACE2 antibody [EPR4435(2)] (ab108252) at 1/1000 dilution
Lane 1 : Wild-type HepG2 cell lysate
Lane 2 : ACE2 knockout HepG2 cell lysate
Lane 3 : Calu-3 cell lysate
Lane 4 : A549 cell lysate
Lysates/proteins at 30 µg per lane.
Performed under reducing conditions.
Predicted band size: 92 kDa
Observed band size: 130 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab108252 observed at 130 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab108252 was shown to react with ACE2 in wild-type HepG2 cells in western blot with loss of signal observed in ACE2 knockout cell line ab273733 (knockout cell lysate ab275495). Wild-type and ACE2 knockout HepG2 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab108252 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACE2 antibody [EPR4435(2)] (ab108252)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labeling ACE2 with ab108252 at 1/6400 dilution. Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. Staining was visualised using Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
The section was incubated with ab108252 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
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All lanes : Anti-ACE2 antibody [EPR4435(2)] (ab108252) at 1/1000 dilution
Lane 1 : Wild-type Caco-2 cell lysate
Lane 2 : ACE2 knockout Caco-2 cell lysate
Lane 3 : Calu-3 cell lysate
Lane 4 : A549 cell lysate
Lysates/proteins at 30 µg per lane.
Performed under reducing conditions.
Predicted band size: 92 kDa
Observed band size: 125 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab108252 observed at 125 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab108252 was shown to react with ACE2 in Caco-2 wild-type cells in western blot with loss of signal observed in ACE2 knockout cell line ab273731 (knockout cell lysate ab275516). Wild-type and ACE2 knockout Caco-2 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab108252 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-ACE2 antibody [EPR4435(2)] (ab108252) at 1/1000 dilution
Lane 1 : Human testis cell lysate
Lane 2 : Human kidney cell lysate
Lane 3 : Human lung cell lysate
Lane 4 : HepG2 cell lysate
Lane 5 : Caco-2 cell lysate
Lane 6 : A549 cell lysate (negative control)
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 92 kDa
Observed band size: 120 kDa why is the actual band size different from the predicted?Lanes 1 - 6: Merged signal (red and green). Green - ab108252 observed at 120 kDa. Red - loading control, Mouse anti-Actin observed at 42kDa.
ab108252 was shown to react with ACE2 in western blot. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab108252 and Mouse anti Actin overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Absence of ACE2 expression in A549 cells aligns with previously reported mRNA and protein data (PMID 16282461; fig.2b and 2c).
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Different batches of ab108252 were tested on Human kidney lysate at 0.2 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 120 kDa.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACE2 antibody [EPR4435(2)] (ab108252)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue labeling ACE2 with ab108252 at 1/6400 dilution. Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. Staining was visualised using Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
The section was incubated with ab108252 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACE2 antibody [EPR4435(2)] (ab108252)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue labeling ACE2 with ab108252 at 1/6400 dilution. Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. Staining was visualised using Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
The section was incubated with ab108252 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
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All lanes : Anti-ACE2 antibody [EPR4435(2)] (ab108252) at 1/1000 dilution
Lane 1 : Human fetal kidney lysate
Lane 2 : Human testis lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 92 kDa -
ab108252 Immunoprecipitating ACE2 in human testis tissue lysate. 0.35 mg of tissue lysate was incubated with 0.6 μg primary antibody (1/20). For western blotting a HRP-conjugated Veriblot for IP Detection Reagent (ab131366) (1/1000) was used to confirm successful immunoprecipitation.
Exposure time: 1 second.
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
All lanes : Anti-ACE2 antibody [EPR4435(2)] (ab108252) at 1/500 dilution
Lane 1 : Human testis tissue lysate at 10 µg
Lane 2 : ab108252 + Human testis tissue lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab108252 in Human testis tissue lysate
Observed band size: 110 kDa why is the actual band size different from the predicted? -
ELISA using ab108252 at varying antibody concentrations (4000~0 ng/ml) and antigen concentration at 1000 ng/mL. An Alkaline Phosphatase-conjugated Goat Anti-Rabbit IgG (H+L) (1/2500) was used as the secondary antibody.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (121)
ab108252 has been referenced in 121 publications.
- Li RJ et al. Qing Fei Hua Xian Decoction ameliorates bleomycin-induced pulmonary fibrosis by suppressing oxidative stress through balancing ACE-AngII-AT1R/ACE2-Ang-(1-7)-Mas axis. Iran J Basic Med Sci 26:107-113 (2023). PubMed: 36594067
- Xia B et al. Extracellular vesicles mediate antibody-resistant transmission of SARS-CoV-2. Cell Discov 9:2 (2023). PubMed: 36609376
- Jidigam VK et al. Histopathological assessments reveal retinal vascular changes, inflammation, and gliosis in patients with lethal COVID-19. Graefes Arch Clin Exp Ophthalmol 260:1275-1288 (2022). PubMed: 34714382
- Villa A et al. DNA aptamers masking angiotensin converting enzyme 2 as an innovative way to treat SARS-CoV-2 pandemic. Pharmacol Res 175:105982 (2022). PubMed: 34798263
- Li D et al. SARS-CoV-2 receptor binding domain radio-probe: a non-invasive approach for angiotensin-converting enzyme 2 mapping in mice. Acta Pharmacol Sin 43:1749-1757 (2022). PubMed: 34815544