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Microbiology Organism Virus RNA Virus ssRNA positive strand virus SARS Coronavirus
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Human ACE2 knockout Caco-2 cell lysate (ab275516)

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Western blot - Human ACE2 knockout Caco-2 cell lysate (ab275516)
  • Western blot - Human ACE2 knockout Caco 2 cell lysate (ab275516)
  • Western blot - Human ACE2 knockout Caco 2 cell lysate (ab275516)
  • Western blot - Human ACE2 knockout Caco 2 cell lysate (ab275516)
  • Sanger Sequencing - Human ACE2 knockout Caco 2 cell lysate

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Anti-ACE2 antibody (ab65863)

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Overview

  • Product name

    Human ACE2 knockout Caco-2 cell lysate
    See all ACE2 kits
  • Product overview


    Knockout cell lysate achieved by CRISPR/Cas9.

  • Parental Cell Line

    Caco 2
  • Organism

    Human
  • Mutation description

    Knockout achieved by using CRISPR/Cas9, Homozygous: 62 bp deletion in exon 2
  • Passage number

    <20
  • Knockout validation

    Sanger Sequencing
  • Reconstitution notes

    To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.

    *Usage of SDS sample buffer is not recommended with these lyophilized lysates.

  • Notes

    Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version - found here. Please refer to our lysis protocol for further details on how our lysates are prepared.

    User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

    Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
    See here for more information on knockout cell lysates.

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

    This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.

  • Tested applications

    Suitable for: WBmore details

Properties

  • Storage instructions

    Store at -80°C. Please refer to protocols.
  • Components 1 kit
    ab277337 - Human ACE2 knockout Caco 2 cell lysate 1 x 100µg
    ab277338 - Human wild-type Caco 2 cell lysate 1 x 100µg
  • Research areas

    • Microbiology
    • Organism
    • Virus
    • RNA Virus
    • ssRNA positive strand virus
    • SARS Coronavirus
    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteolytic enzymes
    • Metalloprotease
    • ACEs
    • Cardiovascular
    • Blood
    • Blood Pressure regulation
    • Cardiovascular
    • Heart
    • Contractility
    • Inotropics
    • Cardiovascular
    • Vasculature
    • Vasoconstriction
    • Other
  • Cell type

    epithelial
  • Disease

    Adenocarcinoma
  • Gender

    Male

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.
  • Target information above from: UniProt accession Q9BYF1 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

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

Associated products

  • KO cell lines

    • Human ACE2 knockout Caco-2 cell line (ab273731)
  • Related Products

    • Anti-ACE2 antibody [EPR4436] (ab108209)
    • Anti-ACE2 antibody [EPR4435(2)] (ab108252)
    • Anti-ACE2 antibody [EPR4435(2)] - BSA and Azide free (ab239924)
    • Anti-ACE2 antibody [EPR4436] - BSA and Azide free (ab247610)
    • Anti-ACE2 antibody (ab65863)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab275516 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
WB
Use at an assay dependent concentration.
Notes
WB
Use at an assay dependent concentration.

Images

  • Western blot - Human ACE2 knockout Caco-2 cell lysate (ab275516)
    Western blot - Human ACE2 knockout Caco-2 cell lysate (ab275516)

    Lane 1: Wild-type Caco-2 cell lysate 30 μg
    Lane 2: ACE2 knockout Caco-2 cell lysate 30 μg
    Lane 3: Calu-3 cell lysate 30 μg
    Lane 4: A549 cell lysate 30 μg
    False colour image of Western blot: Anti-ACE2 antibody staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab15348 was shown to bind specifically to ACE2. A band was observed at 120-135 kDa in wild-type Caco-2 cell lysates with no signal observed at this size in ACE2 knockout cell line ab277338 (knockout cell lysate ab275516). To generate this image, wild-type and ACE2 knockout Caco-2 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

  • Western blot - Human ACE2 knockout Caco 2 cell lysate (ab275516)
    Western blot - Human ACE2 knockout Caco 2 cell lysate (ab275516)

    Lane 1: Wild-type Caco-2 cell lysate 30 ug
    Lane 2: ACE2 knockout Caco-2 cell lysate 30 ug
    Lane 3: Calu-3 cell lysate 30 ug
    Lane 4: A549 cell lysate 30 ug
    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 wild-type Caco-2 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 - Human ACE2 knockout Caco 2 cell lysate (ab275516)
    Western blot - Human ACE2 knockout Caco 2 cell lysate (ab275516)

    Lane 1: Wild-type Caco-2 cell lysate 30 ug
    Lane 2: ACE2 knockout Caco-2 cell lysate 30 ug
    Lane 3: Calu-3 cell lysate 30 ug
    Lane 4: A549 cell lysate 30 ug
    Lanes 1 - 4:Merged signal (red and green). Green - ab108209 observed at 125 kDa. Red - loading controlab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
    ab108209 was shown to react with ACE2 in wild-type Caco-2 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 ab108209 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 - Human ACE2 knockout Caco 2 cell lysate (ab275516)
    Western blot - Human ACE2 knockout Caco 2 cell lysate (ab275516)

    Lane 1: Wild-type Caco-2 cell lysate 30 ug
    Lane 2: ACE2 knockout Caco-2 cell lysate 30 ug
    Lane 3: Calu-3 cell lysate 30 ug
    Lane 4: A549 cell lysate 30 ug
    Lanes 1 - 4:Merged signal (red and green). Green - ab65863 observed at 125 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
    ab65863 was shown to react with ACE2 in wild-type Caco-2 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 ab65863 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at 1 ug/ml 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.

  • Sanger Sequencing - Human ACE2 knockout Caco 2 cell lysate
    Sanger Sequencing - Human ACE2 knockout Caco 2 cell lysate

    Homozygous: 62 bp deletion in exon 2

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

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (0)

Publishing research using ab275516? Please let us know so that we can cite the reference in this datasheet.

ab275516 has not yet been referenced specifically in any publications.

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