Astana Biomed Group, an authorized Abcam distributor in Central Asia

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: ELISA, WB, IP, IHC-P
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-ACE2 antibody [EPR4435(2)]
    See all ACE2 primary antibodies

  • Description

    Rabbit monoclonal [EPR4435(2)] to ACE2

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
    (Peptide available as ab198988)

  • Positive control

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

  • 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
    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.

    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

Images

  • Western blot - Anti-ACE2 antibody [EPR4435(2)] (ab108252)
    Western blot - Anti-ACE2 antibody [EPR4435(2)] (ab108252)
    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACE2 antibody [EPR4435(2)] (ab108252)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACE2 antibody [EPR4435(2)] (ab108252)

    ab108252, at 1/100 dilution, staining ACE2 in paraffin-embedded human kidney tissue by immunohistochemistry.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Western blot - Anti-ACE2 antibody [EPR4435(2)] (ab108252)
    Western blot - Anti-ACE2 antibody [EPR4435(2)] (ab108252)
    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.

  • Western blot - Anti-ACE2 antibody [EPR4435(2)] (ab108252)
    Western blot - Anti-ACE2 antibody [EPR4435(2)] (ab108252)
    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).

  • Western blot - Anti-ACE2 antibody [EPR4435(2)] (ab108252)
    Western blot - Anti-ACE2 antibody [EPR4435(2)] (ab108252)
    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)] (ab108252)
    Immunoprecipitation - Anti-ACE2 antibody [EPR4435(2)] (ab108252)

    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 - Anti-ACE2 antibody [EPR4435(2)] (ab108252)
    ELISA - Anti-ACE2 antibody [EPR4435(2)] (ab108252)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACE2 antibody [EPR4435(2)] (ab108252)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ACE2 antibody [EPR4435(2)] (ab108252)

    ab108252, at 1/100 dilution, staining ACE2 in paraffin-embedded human testis tissue by immunohistochemistry

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Anti-ACE2 antibody [EPR4435(2)] (ab108252)
    Anti-ACE2 antibody [EPR4435(2)] (ab108252)

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