Overview

  • Product name

    Anti-IRAK-1 antibody [EPR19667]
    See all IRAK-1 primary antibodies
  • Description

    Rabbit monoclonal [EPR19667] to IRAK-1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human IRAK-1 aa 200-450. The exact sequence is proprietary.
    Database link: P51617

  • Positive control

    • WB: HeLa, HEK-293, HepG2 and A549 whole cell lysates. ICC/IF: HeLa cells.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab208009 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
WB 1/1000. Detects a band of approximately 80 kDa (predicted molecular weight: 76 kDa).
ICC/IF 1/100.

Target

  • 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
  • Database links

  • Alternative names

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

Images

  • Lane 1: Wild type HAP1 whole cell lysate (40 µg)
    Lane 2: IRAK-1 knockout  HAP1 whole cell lysate (40 µg)
    Lane 3: HeLa whole cell lysate (40 µg)
    Lane 4: HEK293 whole cell lysate (40 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab208009 observed at 85 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab208009 was shown to specifically react with IRAK-1 when IRAK-1 knockout samples were used. Wild-type and IRAK-1 knockout samples were subjected to SDS-PAGE.  Ab208009 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 200 dilution and 1/10000 dilution respectively. Blots were developed with 800CW Goat anti Rabbit and 680CW Goat anti Mouse secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • All lanes : Anti-IRAK-1 antibody [EPR19667] (ab208009) at 1/1000 dilution

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 3 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
    Lane 4 : A549 (Human lung carcinoma cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 76 kDa
    Observed band size: 68,80 kDa
    why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1/3/4: 1 minute; Lane 2: 30 seconds.

    There are two splice variants for IRAK-1, IRAK-1b and IRAK-1c,respectively.   The product recognizes full length IRAK-1 (80 kDa) and IRAK-1c (68 kDa)  (PMID: 16690127).

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling IRAK-1 with ab208009 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing mainly cytoplasmic staining on HeLa cell line. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

    The negative controls are as follows:

    -ve control 1: ab208009 at 1/100 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary at 1/1000 dilution.

    -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary at 1/1000 dilution.

References

This product has been referenced in:

  • Yan M  et al. miR-146b promotes cell proliferation and increases chemosensitivity, but attenuates cell migration and invasion via FBXL10 in ovarian cancer. Cell Death Dis 9:1123 (2018). Read more (PubMed: 30409964) »
See 1 Publication for this product

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