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    products/elisa/human-il-1ra-elisa-kit-ab211650.pdf

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Immunology Innate Immunity Cytokines Interleukins
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Validated using a knockout cell lineRecombinantSimpleStep

Human IL-1ra ELISA Kit (ab211650)

  • Datasheet
  • SDS
  • Protocol Booklet
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Other - Human IL-1ra ELISA Kit (ab211650)
  • Human IL-1RA standard curve comparison data.
  • Example of human IL-1Ra standard curve in Sample Diluent NS.
  • Example of human IL-1Ra standard curve in 1X Cell Extraction Buffer PTR.
  • Interpolated concentrations of native IL-1Ra in human serum and plasma samples.
  • Serum from ten individual healthy human female donors was measured in duplicate.
  • Interpolated concentrations of native IL-1Ra in human saliva, milk and urine samples.
  • Interpolated concentrations of native IL-1Ra in unstimulated and PHA/PMA stimulated human Peripherial Blood Monocyte (PBMC) Cell samples
  • Sandwich ELISA - Human IL-1RA knockout A-431 and THP-1 cells
  • Sandwich ELISA - Human IL-1ra ELISA Kit (ab211650)

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 1.5 pg/ml
  • Range: 7.81 pg/ml - 500 pg/ml
  • Sample type: Cell culture extracts, Cell culture supernatant, Milk, Plasma, Saliva, Serum, Tissue Extracts, Urine
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Human

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Overview

  • Product name

    Human IL-1ra ELISA Kit
    See all IL-1RA kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    EDTA plasma 5 3.5%
    Inter-assay
    Sample n Mean SD CV%
    EDTA plasma 3 4.3%
  • Sample type

    Cell culture supernatant, Saliva, Milk, Urine, Serum, Plasma, Cell culture extracts, Tissue Extracts
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    1.5 pg/ml
  • Range

    7.81 pg/ml - 500 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 98 97% - 99%
    Saliva 102 96% - 112%
    Milk 104 98% - 110%
    Urine 102 100% - 106%
    Serum 106 82% - 119%
    Hep Plasma 103 93% - 117%
    EDTA Plasma 105 100% - 113%
    Cit plasma 94 80% - 102%
  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    Human IL-1ra ELISA Kit (ab211650) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of IL-1ra protein in milk, plasma, saliva, serum, urine, cell culture extracts, tissue extracts, and cell culture supernatant. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human IL-1ra with 4 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.

  • Notes

    Interleukin-1 receptor antagonist (IL-1ra, also known as IL-1F3) is a member of the IL-1 cytokine family. IL-1ra inhibits the activity of interleukin-1 by binding to receptor IL1R1 and preventing its association with the coreceptor IL1RAP for signaling. However, IL-1ra has no interleukin-1 like activity. It binds functional interleukin-1 receptor IL1R1 with greater affinity than decoy receptor IL1R2; however, the physiological relevance of the latter association is unsure.

    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-1Ra Capture Antibody 1 x 600µl
    10X Human IL-1Ra 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 IL-1Ra 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
  • Function

    Inhibits the activity of interleukin-1 by binding to receptor IL1R1 and preventing its association with the coreceptor IL1RAP for signaling. Has no interleukin-1 like activity. Binds functional interleukin-1 receptor IL1R1 with greater affinity than decoy receptor IL1R2; however, the physiological relevance of the latter association is unsure.
  • Tissue specificity

    The intracellular form of IL1RN is predominantly expressed in epithelial cells.
  • Involvement in disease

    Microvascular complications of diabetes 4
    Interleukin 1 receptor antagonist deficiency
  • Cellular localization

    Cytoplasm and Secreted.
  • Target information above from: UniProt accession P18510 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • DIRA
    • F630041P17Rik
    • ICIL 1RA
    • ICIL-1RA
    • ICIL1RA
    • IL-1ra
    • IL-1ra3
    • IL-1RN
    • IL1 inhibitor
    • IL1F3
    • IL1RA
    • IL1RA_HUMAN
    • IL1RN
    • IL1RN (IL1F3)
    • Interleukin 1 receptor antagonist
    • Interleukin-1 receptor antagonist protein
    • Intracellular IL 1 receptor antagonist type II
    • Intracellular interleukin 1 receptor antagonist (icIL 1ra)
    • IRAP
    • MGC10430
    • MVCD4
    • Type II interleukin 1 receptor antagonist
    see all
  • Database links

    • Entrez Gene: 3557 Human
    • Omim: 147679 Human
    • SwissProt: P18510 Human
    • Unigene: 81134 Human

    Associated products

    • Alternative Versions

      • Human IL-1ra Antibody Pair - BSA and Azide free (ab241800)
    • KO cell lines

      • Human IL1RN (IL1 Receptor Antagonist) knockout A-431 cell line (ab273379)

    Images

    • Other - Human IL-1ra ELISA Kit (ab211650)
      Other - Human IL-1ra ELISA Kit (ab211650)

      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.

       

    • Human IL-1RA standard curve comparison data.
      Human IL-1RA standard curve comparison data.

      Standard curve comparison between human IL-1RA SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows a 4-fold increase in sensitivity.

    • Example of human IL-1Ra standard curve in Sample Diluent NS.
      Example of human IL-1Ra standard curve in Sample Diluent NS.

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

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

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

    • Interpolated concentrations of native IL-1Ra in human serum and plasma samples.
      Interpolated concentrations of native IL-1Ra in human serum and plasma samples.

      The concentrations of IL-1Ra were measured in duplicates, interpolated from the IL-1Ra standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 100%, plasma (EDTA) 100%, plasma (heparin) 100%; and plasma (citrate) 100%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IL-1Ra concentration was determined to be 276 pg/mL in serum, 239 pg/mL in plasma (EDTA) and 233 pg/mL in plasma (heparin) and 170 pg/mL in plasma (citrate).

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

      Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IL-1Ra concentration was determined to be 277 pg/mL with a range of 130 – 598 pg/mL.

    • Interpolated concentrations of native IL-1Ra in human saliva, milk and urine samples.
      Interpolated concentrations of native IL-1Ra in human saliva, milk and urine samples.

      The concentrations of IL-1Ra were measured in duplicates, interpolated from the IL-1Ra standard curves and corrected for sample dilution. Undiluted samples are as follows: saliva 1%, milk 15%, and urine 15%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IL-1Ra concentration was determined to be 207 ng/mL in saliva, 1,121 pg/mL in milk and 4,271 pg/mL in urine.

    • Interpolated concentrations of native IL-1Ra in unstimulated and PHA/PMA stimulated human Peripherial Blood Monocyte (PBMC) Cell samples
      Interpolated concentrations of native IL-1Ra in unstimulated and PHA/PMA stimulated human Peripherial Blood Monocyte (PBMC) Cell samples

      The concentrations of IL-1Ra were measured in duplicates, interpolated from the IL-1Ra standard curves and corrected for sample dilution. Undiluted samples are as follows: unstimulated 2% and stimulated 2%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IL-1Ra concentration was determined to be 3360 pg/mL in unstimulated, 11,912 pg/mL in stimulated and undetectable in media.

    • Sandwich ELISA - Human IL-1RA knockout A-431 and THP-1 cells
      Sandwich ELISA - Human IL-1RA knockout A-431 and THP-1 cells

      Human IL-1RA concentration was interpolated from the IL-1RA standard curve. Supernatants from cell culture samples were serially diluted and assessed by the Human IL-1ra ELISA Kit (ab211650). Wild-type A-431, IL1RN knockout A-431 (ab273379) and THP-1 cells were assessed in duplicate (n=2). Data are represented as the mean and error bars represent standard deviation.

    • Sandwich ELISA - Human IL-1ra ELISA Kit (ab211650)
      Sandwich ELISA - Human IL-1ra ELISA Kit (ab211650)
      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 (2)

    Publishing research using ab211650? Please let us know so that we can cite the reference in this datasheet.

    ab211650 has been referenced in 2 publications.

    • Kuemmerle-Deschner JB  et al. New variant in the IL1RN-gene (DIRA) associated with late-onset, CRMO-like presentation. Rheumatology (Oxford) 59:3259-3263 (2020). PubMed: 32259833
    • Karpenko MN  et al. Interleukin-1ß, interleukin-1 receptor antagonist, interleukin-6, interleukin-10, and tumor necrosis factor-a levels in CSF and serum in relation to the clinical diversity of Parkinson's disease. Cell Immunol 327:77-82 (2018). ELISA ; Human . PubMed: 29478949

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