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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)

Reviews (2) Submit a question References (5)
Western blot - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)
  • Western blot - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)
  • Western blot - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)
  • Western blot - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)
  • Immunoprecipitation - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)
  • Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)

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 - BSA and Azide free
  • Suitable for: Indirect ELISA, IP, IHC-P, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Unconjugated

Overview

  • Product name

    Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free
    See all ACE2 primary antibodies

  • Description

    Rabbit monoclonal [EPR4435(2)] to ACE2 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Indirect ELISA, IP, IHC-P, WBmore 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 as ab198988)

  • Positive control

    • IHC-P: Human, mouse, and rat kidney tissues; WB: Caco-2, HepG2 and Calu-3 cell lysates; Human fetal kidney and human testis lysates; IP: Human testis tissue lysate.

  • General notes

    ab239924 is the carrier-free version of ab108252.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab239924 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
Indirect ELISA
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB (1)
Use at an assay dependent concentration. Predicted molecular weight: 92 kDa.Can be blocked with ACE2 peptide (ab198988).
Notes
Indirect ELISA
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB
Use at an assay dependent concentration. Predicted molecular weight: 92 kDa.Can be blocked with ACE2 peptide (ab198988).
Application notes
Is unsuitable for Flow Cyt or ICC/IF.

Target

  • 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
    modifications

    N-glycosylation on Asn-90 may limit SARS infectivity.

  • Cellular localization

    Secreted and Cell membrane.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • ACE 2 antibody
    • ACE related carboxypeptidase antibody
    • ACE-related carboxypeptidase antibody
    • ACE2 antibody
    • ACE2_HUMAN antibody
    • ACEH antibody
    • Angiotensin converting enzyme 2 antibody
    • Angiotensin converting enzyme homolog antibody
    • Angiotensin converting enzyme like protein antibody
    • Angiotensin I Converting Enzyme (peptidyl dipeptidase A) 2 antibody
    • Angiotensin I converting enzyme 2 antibody
    • Angiotensin-converting enzyme homolog antibody
    • DKFZP434A014 antibody
    • EC 3.4.17 antibody
    • metalloprotease MPROT 15 antibody
    • Metalloprotease MPROT15 antibody
    • OTTHUMP00000022963 antibody
    • Processed angiotensin-converting enzyme 2 antibody
    see all

Images

  • Western blot - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)
    Western blot - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)
    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?



    This data was developed using ab108252, the same antibody clone in a different buffer formulation.

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)

    This data was developed using ab108252, the same antibody clone in a different buffer formulation.

    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.

  • Western blot - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)
    Western blot - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)
    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?



    This data was developed using the same antibody clone in a different buffer formulation (ab108252).

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)

    This data was developed using ab108252, the same antibody clone in a different buffer formulation.

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)

    This data was developed using ab108252, the same antibody clone in a different buffer formulation.

    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.

  • Western blot - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)
    Western blot - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)
    This data was developed using ab108252, the same antibody clone in a different buffer formulation. 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.
  • Western blot - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)
    Western blot - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)
    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

  • Immunoprecipitation - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)
    Immunoprecipitation - Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)

    ab239924 Immunoprecipitating ACE2 in human testis tissue lysate. 0.35 mg of tissue lysate was incubated with 2 μg primary antibody (1/50). 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)] - BSA and Azide free (ab239924) at 1/500 dilution

    Lane 1 : Human testis tissue lysate at 10 µg
    Lane 2 : ab239924 + Human testis tissue lysate
    Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab239924 in Human testis tissue lysate

    Observed band size: 110 kDa why is the actual band size different from the predicted?

  • Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)
    Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (5)

Publishing research using ab239924? Please let us know so that we can cite the reference in this datasheet.

ab239924 has been referenced in 5 publications.

  • Maccio U  et al. SARS-CoV-2 leads to a small vessel endotheliitis in the heart. EBioMedicine 63:103182 (2021). PubMed: 33422990
  • Lee IT  et al. Robust ACE2 protein expression localizes to the motile cilia of the respiratory tract epithelia and is not increased by ACE inhibitors or angiotensin receptor blockers. medRxiv N/A:N/A (2020). PubMed: 32511516
  • Dobrindt K  et al. Common genetic variation in humans impacts in vitro susceptibility to SARS-CoV-2 infection. bioRxiv N/A:N/A (2020). PubMed: 32995783
  • Blanco-Melo D  et al. Imbalanced Host Response to SARS-CoV-2 Drives Development of COVID-19. Cell 181:1036-1045.e9 (2020). PubMed: 32416070
  • Lee IT  et al. ACE2 localizes to the respiratory cilia and is not increased by ACE inhibitors or ARBs. Nat Commun 11:5453 (2020). PubMed: 33116139

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Western blot abreview for Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free

 Good
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mice hepatocytes)
Gel Running Conditions
Reduced Denaturing (4-15 % SDS-PAGE)
Loading amount
35 µg
Specification
Mice hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Aug 19 2020

Immunocytochemistry/ Immunofluorescence abreview for Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free

 Good
Abreviews
This product is known to not work in this application or species.
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Mice hepatocytes)
Permeabilization
Yes - 0.2% Triton X-100 in PBS
Specification
Mice hepatocytes
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Armen Petrosyan

Verified customer

Submitted Aug 03 2020

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