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  1. Link

    human-ace2-elisa-kit-ab235649.pdf

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Microbiology Organism Virus RNA Virus ssRNA positive strand virus SARS Coronavirus
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RecombinantSimpleStep

Human ACE2 ELISA Kit (ab235649)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (1) Submit a question References (7)

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SimpleStep ELISA Protocol Diagram
  • Example of human ACE2 standard curve in 1X Cell Extraction Buffer PTR.
  • Interpolated concentrations of native ACE2 in human urine samples.
  • Interpolated concentrations of native ACE2 in human lung, heart, fetal intestine, kidney, and testes samples.
  • Sandwich ELISA - Human ACE2 ELISA Kit (ab235649)

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 1052 pg/ml
  • Range: 3.98 ng/ml - 255 ng/ml
  • Sample type: Cell culture media, Cit plasma, EDTA Plasma, Hep Plasma, Serum, Tissue Extracts, Urine
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Human

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Overview

  • Product name

    Human ACE2 ELISA Kit
    See all ACE2 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    extract 8 2.3%
    Inter-assay
    Sample n Mean SD CV%
    extract 3 3.2%
  • Sample type

    Urine, Serum, Tissue Extracts, Cell culture media, Hep Plasma, EDTA Plasma, Cit plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    1052 pg/ml
  • Range

    3.98 ng/ml - 255 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Urine 88 85% - 89%
    Serum 99 % - %
    Tissue Extracts 104 100% - 108%
    Cell culture media 112 110% - 116%
    Hep Plasma 82 81% - 84%
    EDTA Plasma 91 90% - 92%
    Cit plasma 99 98% - 100%
  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Human ACE2 ELISA Kit (ab235649) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of native ACE2 protein in tissue extracts and urine. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human ACE2 with 1052 pg/ml sensitivity.


    Note: Plasma, serum, and cell culture media samples were validated with spiked recombinant ACE2. Neat serum from ten individual healthy human female/male donors was measured in duplicate and all values were below the detectable range of the assay.


    SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:


            - Single-wash protocol reduces assay time to 90 minutes or less
            - High sensitivity, specificity and reproducibility from superior antibodies
            - Fully validated in biological samples
            - 96-wells plate breakable into 12 x 8 wells strips


    A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.

  • Notes

    ACE2 is a key enzyme in the renin-angiotensin system which regulates blood pressure. Specifically, ACE2 cleaves the terminal nonapeptide from angiotensin I or the terminal septapeptide from angiotensin II to create angiotensin[1-9] and angiotensin[1-7], respectively. The produced peptides work to oppose the effects of angiotensin II and act as a vasodilator. This enzyme is currently under investigation as a potential target for reducing instances of heart failure.

    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.

  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Human ACE2 Capture Antibody 1 x 600µl
    10X Human ACE2 Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent 4BI 1 x 6ml
    Human ACE2 Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 12ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • 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
  • 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
  • Database links

    • Entrez Gene: 59272 Human
    • Omim: 300335 Human
    • SwissProt: Q9BYF1 Human
    • Unigene: 178098 Human

    Associated products

    • Alternative Versions

      • Human ACE2 Antibody Pair - BSA and Azide free (ab244135)

    Images

    • SimpleStep ELISA Protocol Diagram
      SimpleStep ELISA Protocol Diagram

      SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

       

    • Example of human ACE2 standard curve in 1X Cell Extraction Buffer PTR.
      Example of human ACE2 standard curve in 1X Cell Extraction Buffer PTR.

      Background-subtracted data values (mean +/- SD) are graphed.

    • Interpolated concentrations of native ACE2 in human urine samples.
      Interpolated concentrations of native ACE2 in human urine samples.

      The concentrations of ACE2 were measured in duplicates, interpolated from the ACE2 standard curves and corrected for sample dilution. Undiluted samples are as follows: urine 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean ACE2 concentration was determined to be 75.24 ng/mL in urine.

    • Interpolated concentrations of native ACE2 in human lung, heart, fetal intestine, kidney, and testes samples.
      Interpolated concentrations of native ACE2 in human lung, heart, fetal intestine, kidney, and testes samples.

      The concentrations of ACE2 were measured in duplicate and interpolated from the ACE2 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean ACE2 concentration was determined to be 26 ng/mL in lung extract, 31 ng/mL in heart extract, 115 ng/mL in fetal intestine, 94 ng/mL in kidney, and 146 ng/mL in testes extract.

    • Sandwich ELISA - Human ACE2 ELISA Kit (ab235649)
      Sandwich ELISA - Human ACE2 ELISA Kit (ab235649)
      To learn more about the advantages of recombinant antibodies see here.

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (7)

    Publishing research using ab235649? Please let us know so that we can cite the reference in this datasheet.

    ab235649 has been referenced in 7 publications.

    • Takahashi Y  et al. Histone deacetylase inhibitors suppress ACE2 and ABO simultaneously, suggesting a preventive potential against COVID-19. Sci Rep 11:3379 (2021). PubMed: 33564039
    • Wang C  et al. Membrane Nanoparticles Derived from ACE2-Rich Cells Block SARS-CoV-2 Infection. ACS Nano 15:6340-6351 (2021). PubMed: 33734675
    • Maggi F  et al. Nicotine upregulates ACE2 expression and increases competence for SARS-CoV-2 in human pneumocytes. ERJ Open Res 7:N/A (2021). PubMed: 33850935
    • Higuchi Y  et al. Engineered ACE2 receptor therapy overcomes mutational escape of SARS-CoV-2. Nat Commun 12:3802 (2021). PubMed: 34155214
    • Russo P  et al. COVID-19 and smoking: is nicotine the hidden link? Eur Respir J 55:N/A (2020). PubMed: 32341101
    • Li S  et al. Exploring the Demographics and Clinical Characteristics Related to the Expression of Angiotensin-Converting Enzyme 2, a Receptor of SARS-CoV-2. Front Med (Lausanne) 7:530 (2020). PubMed: 32974373
    • Cocozza F  et al. Extracellular vesicles containing ACE2 efficiently prevent infection by SARS-CoV-2 Spike protein-containing virus. J Extracell Vesicles 10:e12050 (2020). PubMed: 33391636

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Human ACE2 measurement in cell lysates

    Excellent Excellent 5/5 (Ease of Use)
    Abreviews
    Abreviews
    abreview image
    We used the ab235649 Human ACE2 SimpleStep ELISA kit to analyze ACE2 levels in cell lysate of bronchial epithelial Calu-3 cells to verify if our stimulus influences ACE2 expression.
    In order to measure ACE-2 levels in the cell lysate we modified the existing indications for tissue extract. After treatment, Calu-3 cell monolayer was washed with 200 µL PBS to remove any residue of cell medium and 120 µL of 1X Cell extraction buffer PTR was added. With the help of a scraper and by pipetting up and down, cells were lysated and collected in a 1.5 mL tube. Samples were incubated on ice for 20 minutes and then centrifuged at 13000 rpm for 10 min at 4°C. The supernatant obtained was transferred into clean tubes and analysed. Samples were diluted 1:10 in 1X Cell extraction buffer PTR to be analysed.
    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 May 27 2022

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