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  1. Link

    human-il-18-elisa-kit-ab215539.pdf

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Immunology Innate Immunity Cytokines Interleukins
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RecombinantSimpleStep

Human IL-18 ELISA Kit (ab215539)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (1) Submit a question References (4)

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Other - Human IL-18 ELISA Kit (ab215539)
  • Example of human IL-18 standard curve in Sample Diluent NS.
  • Example of human IL-18 standard curve in Sample Diluent NS.
  • Example of human IL-18 standard curve in 1X Cell Extraction Buffer PTR.
  • Example of human IL-18 standard curve in 1X Cell Extraction Buffer PTR.
  • Interpolated concentrations of native IL-18 in human HeLa, HL-60, and A431 Cell Extract samples based on a 800 µg/ml extract load.
  • Interpolated concentrations of spike IL-18 in human serum and plasma samples.
  • Serum from nine individual healthy human female donors was measured in duplicate.
  • Interpolated concentrations of spike IL-18 in cell culture media and human urine samples.
  • Linearity of dilution.
  • Linearity of dilution.
  • Assay sensitivity.
  • Sandwich ELISA - Human IL-18 ELISA Kit (ab215539)

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 8.3 pg/ml
  • Range: 62.5 pg/ml - 4000 pg/ml
  • Sample type: Cell culture extracts, Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Serum, Tissue Extracts, Urine
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Human

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Protein
Recombinant Human IL-18 protein (ab188059)
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Overview

  • Product name

    Human IL-18 ELISA Kit
    See all IL-18 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    HeLa extract 5 2.7%
    Inter-assay
    Sample n Mean SD CV%
    HeLa extract 3 14.2%
  • Sample type

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

    Sandwich (quantitative)
  • Sensitivity

    8.3 pg/ml
  • Range

    62.5 pg/ml - 4000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Urine 88 86% - 91%
    Serum 84 80% - 91%
    Cell culture extracts 96 93% - 101%
    Cell culture media 96 89% - 104%
    Hep Plasma 93 91% - 97%
    EDTA Plasma 86 82% - 91%
    Cit plasma 82 80% - 84%
  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
    Does not react with: Cow
  • Product overview

    Human IL-18 ELISA Kit (ab215539) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of IL-18 protein in cell culture extracts, cell culture supernatant, cit plasma, edta plasma, hep plasma, serum, tissue extracts, and urine. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human IL-18 with 8.3 pg/ml sensitivity.


    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.


    ASSAY SPECIFICITY  This kit recognizes both native and recombinant human IL-18 protein in serum, plasma, and cell culture supernatant, cell and tissue extract samples only.


    CROSS REACTIVITY Recombinant IL-18BP was prepared at 50 ng/mL and 8 ng/mL and assayed for cross reactivity. No cross-reactivity was observed.


    INTERFERENCE Recombinant IL-18BP was prepared at 50 ng/mL and 8 ng/mL and tested for interference. No interference with was observed.


    SPECIES REACTIVITY This kit recognizes human IL-18 protein.
    Other species reactivity was determined by measuring 50 % serum samples of various species, interpolating the protein concentrations from the human standard curve, and expressing the interpolated concentrations as a percentage of the protein concentration in human serum assayed at the same dilution.
    Reactivity < 3% was determined for the following species: Mouse, rat and cow.

  • Notes

    Interleukin-18 (IL-18 or Interferon gamma-inducing factor) is a proinflammatory cytokine in the IL-1 superfamily. IL-18 has overlapping and synergistic biological function with IL-12. IL-18 is found in both intracellular and extracellular compartments. The antibodies in this kit were raised to the mature full-length sequence of human IL-18. Human IL-18 has 66% protein sequence identity to mouse Lipocalin-2.

    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 IL-18 Capture Antibody 1 x 600µl
    10X Human IL-18 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 CPI - HAMA Blocker (ab193969) 1 x 6ml
    Human IL-18 Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

    • Immunology
    • Innate Immunity
    • Cytokines
    • Interleukins
    • Cancer
    • Invasion/microenvironment
    • Angiogenesis
    • Angiogenic growth factors
    • Cancer
    • Tumor immunology
    • Cytokines
    • Interleukins
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia
  • Function

    Augments natural killer cell activity in spleen cells and stimulates interferon gamma production in T-helper type I cells.
  • Sequence similarities

    Belongs to the IL-1 family.
  • Cellular localization

    Secreted.
  • Target information above from: UniProt accession Q14116 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • Iboctadekin
    • IFN gamma inducing factor
    • IFN-gamma-inducing factor
    • IGIF
    • IL 1 gamma
    • IL 18
    • IL 1g
    • IL-1 gamma
    • IL-18
    • IL1 gamma
    • IL18
    • IL18 protein
    • IL18_HUMAN
    • IL1F4
    • IL1g
    • IL1gamma
    • ILIF4
    • Interferon gamma inducing factor
    • Interferon gamma-inducing factor
    • Interleukin 1 gamma
    • Interleukin 18
    • Interleukin 18 (interferon-gamma-inducing factor)
    • Interleukin-1 gamma
    • Interleukin-18
    • Interleukin18
    • MGC12320
    see all
  • Database links

    • Entrez Gene: 3606 Human
    • Omim: 600953 Human
    • SwissProt: Q14116 Human
    • Unigene: 83077 Human

    Associated products

    • ELISA kits

      • Human IL-18 Matched Antibody Pair Kit (ab218185)

    Images

    • Other - Human IL-18 ELISA Kit (ab215539)
      Other - Human IL-18 ELISA Kit (ab215539)

      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 IL-18 standard curve in Sample Diluent NS.
      Example of human IL-18 standard curve in Sample Diluent NS.

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

    • Example of human IL-18 standard curve in Sample Diluent NS.
      Example of human IL-18 standard curve in Sample Diluent NS.

      Example of human IL-18 standard curve in Sample Diluent NS. The IL-18 standard curve was prepared as described. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

    • Example of human IL-18 standard curve in 1X Cell Extraction Buffer PTR.
      Example of human IL-18 standard curve in 1X Cell Extraction Buffer PTR.

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

    • Example of human IL-18 standard curve in 1X Cell Extraction Buffer PTR.
      Example of human IL-18 standard curve in 1X Cell Extraction Buffer PTR.

      Example of human IL-18 standard curve in 1X Cell Extraction Buffer PTR. The IL-18 standard curve was prepared as described. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

    • Interpolated concentrations of native IL-18 in human HeLa, HL-60, and A431 Cell Extract samples based on a 800 µg/ml extract load.
      Interpolated concentrations of native IL-18 in human HeLa, HL-60, and A431 Cell Extract samples based on a 800 µg/ml extract load.

      The concentrations of IL-18 were measured in duplicate and interpolated from the IL-18 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IL-18 concentration was determined to be 1,099 pg/ml in HeLa cell extract, 456 pg/ml in HL-60 cell extract and 567 pg/ml in A431 cell extract.

    • Interpolated concentrations of spike IL-18 in human serum and plasma samples.
      Interpolated concentrations of spike IL-18 in human serum and plasma samples.

      The concentrations of IL-18 were measured in duplicates, interpolated from the IL-18 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 50%, plasma (citrate) 50%, plasma (EDTA), and plasma (heparin) 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IL-18 concentration was determined to be 3,681 pg/mL in serum, 3,692 pg/mL in plasma (citrate), 6,941pg/mL in plasma (EDTA) and 4,057 pg/mL in plasma (heparin).

    • Serum from nine individual healthy human female donors was measured in duplicate.
      Serum from nine individual healthy human female donors was measured in duplicate.

      Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IL-18 concentration was determined to be 423 pg/mL with a range of undetectable – 2,852 pg/mL.

    • Interpolated concentrations of spike IL-18 in cell culture media and human urine samples.
      Interpolated concentrations of spike IL-18 in cell culture media and human urine samples.

      The concentrations of IL-18 were measured in duplicates, interpolated from the IL-18 standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50% and urine 12.5%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IL-18 concentration was determined to be 3,079 pg/mL in cell culture media and 4,119 pg/mL in urine.

    • Linearity of dilution.
      Linearity of dilution.

      Linearity of dilution is determined based on interpolated values from the standard curve. Linearity of dilution defines a sample concentration interval in which interpolated target concentrations are directly proportional to sample dilution.

      Native IL-18 was measured in the following biological samples in a 2-fold dilution series. Sample dilutions are made in 1X Cell Extraction Buffer PTR.

    • Linearity of dilution.
      Linearity of dilution.

      Recombinant IL-18 was spiked into the following biological samples and diluted in a 2-fold dilution series in Sample Diluent NS.

    • Assay sensitivity.
      Assay sensitivity.

      The MDD was determined by calculating the mean of zero standard replicates and adding 2 standard deviations then extrapolating the corresponding concentration.

    • Sandwich ELISA - Human IL-18 ELISA Kit (ab215539)
      Sandwich ELISA - Human IL-18 ELISA Kit (ab215539)
      To learn more about the advantages of recombinant antibodies see here.

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet
    • SDS
  • References (4)

    Publishing research using ab215539? Please let us know so that we can cite the reference in this datasheet.

    ab215539 has been referenced in 4 publications.

    • Ma C  et al. LncRNA GAS5 overexpression downregulates IL-18 and induces the apoptosis of fibroblast-like synoviocytes. Clin Rheumatol N/A:N/A (2019). PubMed: 31372853
    • Zhong F  et al. miR-145 eliminates lipopolysaccharides-induced inflammatory injury in human fibroblast-like synoviocyte MH7A cells. J Cell Biochem 119:10059-10066 (2018). PubMed: 30191608
    • Song E  et al. Targeted proteomic assays for quantitation of proteins identified by proteogenomic analysis of ovarian cancer. Sci Data 4:170091 (2017). PubMed: 28722704
    • Wu J  et al. Urinary RBP and NGAL Levels are Associated with Nephropathy in Patients with Type 2 Diabetes. Cell Physiol Biochem 42:594-602 (2017). PubMed: 28954270

    Customer reviews and Q&As

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    SimpleStep ELISA saved lots of time

    Excellent Excellent 5/5 (Ease of Use)
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    I've used multiple ELISA kit but among all of them this SimpleStep ELISA kit has the easiest, simplest and shortest procedure. The results are also reliable and standard graph turned out very nicely.

    Abcam user community

    Verified customer

    Submitted Jan 23 2018

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