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RabMAb

Anti-IRAK1抗体[EPR18630] (ab180747)

概述

  • 产品名称
    Anti-IRAK1抗体[EPR18630]
    参阅全部 IRAK1 一抗
  • 描述
    兔单克隆抗体[EPR18630] to IRAK1
  • 宿主
    Rabbit
  • 经测试应用
    适用于: WB, ICC/IF, IPmore details
  • 种属反应性
    与反应: Human
  • 免疫原

    Recombinant fragment within Human IRAK aa 200-500. The exact sequence is proprietary.
    Database link: P51617

  • 阳性对照
    • WB: HEK-293, Jurkat, K562 and HeLa whole cell lysates. ICC/IF: HeLa and K562 cells. IP: HEK-293 whole cell lysate.
  • 常规说明

     

     

    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.

性能

应用

Our Abpromise guarantee covers the use of ab180747 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB 1/1000. Detects a band of approximately 77 kDa (predicted molecular weight: 77 kDa).
ICC/IF 1/250.
IP 1/100.

靶标

  • 功能
    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.
  • 组织特异性
    Isoform 1 and isoform 2 are ubiquitously expressed in all tissues examined, with isoform 1 being more strongly expressed than isoform 2.
  • 序列相似性
    Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family. Pelle subfamily.
    Contains 1 protein kinase domain.
  • 翻译后修饰
    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
  • 数据库链接
  • 别名
    • 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

图片

  • Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: IRAK1 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: Hek293 whole cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab180747 observed at 85 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab180747 was shown to specifically react with IRAK1 when IRAK1 knockout samples were used. Wild-type and IRAK1 knockout samples were subjected to SDS-PAGE. Ab180747 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 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-IRAK1 antibody [EPR18630] (ab180747) at 1/1000 dilution

    Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 2 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 3 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate
    Lane 4 : HeLa (Human epithelial cell line from cervix adenocarcinoma) 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: 77 kDa
    Observed band size: 77 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 10 seconds; Lane 2: 3 minutes; Lane 3: 30 seconds; Lane 4: 1 minute.

    There are two splice variants for IRAK1, IRAK1b and IRAK1c, respectively.  The product recognizes full length IRAK1 (80kDa).

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling IRAK1 with ab180747 at 1/250 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HeLa cells. 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: ab180747 at 1/250 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.
    -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cell line from bone marrow) cells labeling IRAK1 with ab180747 at 1/250 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on K562 cells. 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: ab180747 at 1/250 dilution, followed by followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.
    -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • IRAK1 was immunoprecipitated from 0.35 mg of HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate with ab180747 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab180747 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/1000 dilution.

    Lane 1: HEK-293 whole cell lysate 10µg (Input).

    Lane 2: ab180747 IP in HEK-293 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab180747 in HEK-293 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 30 seconds.

    The 68kDa band observed is likely a splice variant, according to PMID: 16690127.

文献

ab180747 has not yet been referenced specifically in any publications.

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