Overview

  • Product name

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

    Fluorescent
  • Precision

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

    Cell culture supernatant, Saliva, Milk, Urine, Serum, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    1.4 pg/ml
  • Range

    3.9 pg/ml - 2000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Saliva 102 96% - 112%
    Milk 104 98% - 110%
    Urine 102 100% - 106%
    Serum 106 82% - 119%
    Plasma 103 93% - 117%
    Cell culture media 98 97% - 99%
    EDTA Plasma 105 100% - 113%
    Citrate Plasma 94 80% - 102%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse, Rat
  • Product overview

    IL-1Ra in vitro CatchPoint SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of IL-1Ra protein in human serum, plasma, cell culture supernatants, milk, saliva, and urine.


    This CatchPoint SimpleStep ELISA kit has been optimized for Molecular Devices Microplate Readers. Click here for a list of recommended Microplate Readers.
    If using a Molecular Devices’ plate reader supported by SoftMax® Pro software, a preconfigured protocol for these CatchPoint SimpleStep ELISA Kits is available with all the protocol and analysis settings at www.softmaxpro.org.


    The CatchPoint SimpleStep ELISA employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material.  CatchPoint HRP Development Solution containing the Stoplight Red Substrate is added. During incubation, the substrate is catalyzed by HRP generating a fluorescent product. Signal is generated proportionally to the amount of bound analyte and the intensity is measured in a fluorescence plater reader at 530/570/590 nm Excitation/Cutoff/Emission.

  • 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.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • 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
    100X Stoplight Red Substrate 1 x 120µl
    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
    500X Hydrogen Peroxide (H2O2, 3%) 1 x 50µl
    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 1 x 50ml
    SimpleStep Pre-Coated Black 96-Well Microplate 1 unit
    Stoplight Red Substrate Buffer 1 x 12ml
  • Research areas

  • 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.
  • 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

Applications

Our Abpromise guarantee covers the use of ab229435 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • 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.

     

  • The IL-1Ra standard curve was prepared as described in Section 10.

  • 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).

  • 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.

  • 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.

  • 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.

Protocols

References

ab229435 has not yet been referenced specifically in any publications.

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