Overview

  • Product name

    Human IRAK-1 ELISA Kit
    See all IRAK-1 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 5 3.3%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 4%
  • Sample type

    Cell culture extracts
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    22 pg/ml
  • Range

    156 pg/ml - 10000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture extracts 110 107% - 113%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    IRAK-1 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of IRAK-1 protein in human cell extracts.


    The 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. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Notes

    IRAK-1 is a serine/threonine-protein kinase involved in the innate immune response against pathogens. IRAK-1’s association with MYC88 after TLR activation leads to its phosphorylation by IRAK-4. IRAK-1 phosphorylation eventually results in the transcriptional activation of type 1 IFN genes.

  • 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
    10X Human IRAK-1 Capture Antibody 1 x 600µl
    10X Human IRAK-1 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 IRAK-1 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

  • Function

    Binds to the IL-1 type I receptor following IL-1 engagement, triggering intracellular signaling cascades leading to transcriptional up-regulation and mRNA stabilization. Isoform 1 binds rapidly but is then degraded allowing isoform 2 to mediate a slower, more sustained response to the cytokine. Isoform 2 is inactive suggesting that the kinase activity of this enzyme is not required for IL-1 signaling. Once phosphorylated, IRAK1 recruits the adapter protein PELI1.
  • Tissue specificity

    Isoform 1 and isoform 2 are ubiquitously expressed in all tissues examined, with isoform 1 being more strongly expressed than isoform 2.
  • Sequence similarities

    Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family. Pelle subfamily.
    Contains 1 protein kinase domain.
  • Post-translational
    modifications

    Autophosphorylated or is transphosphorylated by IRAK4 following recruitment to the IL-1RI. In the case of isoform 1, this is linked to ubiquitination and degradation.
    Polyubiquitinated; after cell stimulation with IL-1-beta. Polyubiquitination occurs with polyubiquitin chains linked through 'Lys-63'.
  • Information by UniProt
  • Alternative names

    • AA48924
    • Il1rak
    • Interleukin 1 receptor associated kinase 1
    • Interleukin-1 receptor-associated kinase 1
    • IRAK
    • IRAK-1
    • Irak1
    • IRAK1-S
    • IRAK1_HUMAN
    • mPLK
    • OTTHUMP00000026014
    • OTTHUMP00000026015
    • OTTHUMP00000026020
    • OTTHUMP00000180621
    • Pelle
    • Pelle homolog
    • Pelle-like protein kinase
    • Plpk
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab219630 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.

     

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

  • The concentrations of IRAK-1 were measured in duplicate and interpolated from the IRAK-1 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean IRAK-1 concentration was determined to be 8178 pg/mL in MCF7 extracts, 3768 pg/mL in HEK93T extracts, and 3927 pg/mL in K562 extracts.

Protocols

References

ab219630 has not yet been referenced specifically in any publications.

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