製品の概要

  • 製品名
  • 製品の詳細
    Rabbit polyclonal to IKB alpha
  • アプリケーション
    適用あり: WB, ICC/IFmore details
  • 種交差性
    交差種: Human
    交差が予測される動物種: Mouse, Rat, Sheep, Cow, Pig
  • 免疫原

    Synthetic peptide corresponding to Human IKB alpha aa 1-100 conjugated to Keyhole Limpet Haemocyanin (KLH).
    Database link: P25963
    (Peptide available as ab105485)

  • ポジティブ・コントロール
    • This antibody gave a positive signal in HAP1 (WT) and HeLa whole cell lysates, Human kidney and Human spleen tissue lysates. ICC/IF: HeLa cells.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab95338 in the following tested applications.

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

アプリケーション Abreviews 特記事項
WB Use a concentration of 1 µg/ml. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).
ICC/IF Use a concentration of 1 µg/ml.

ターゲット情報

  • 機能
    Inhibits the activity of dimeric NF-kappa-B/REL complexes by trapping REL dimers in the cytoplasm through masking of their nuclear localization signals. On cellular stimulation by immune and proinflammatory responses, becomes phosphorylated promoting ubiquitination and degradation, enabling the dimeric RELA to translocate to the nucleus and activate transcription.
  • 関連疾患
    Ectodermal dysplasia, anhidrotic, with T-cell immunodeficiency autosomal dominant
  • 配列類似性
    Belongs to the NF-kappa-B inhibitor family.
    Contains 5 ANK repeats.
  • 翻訳後修飾
    Phosphorylated; disables inhibition of NF-kappa-B DNA-binding activity. Phosphorylation at positions 32 and 36 is prerequisite to recognition by UBE2D3 leading to polyubiquitination and subsequent degradation.
    Sumoylated; sumoylation requires the presence of the nuclear import signal. Sumoylation blocks ubiquitination and proteasome-mediated degradation of the protein thereby increasing the protein stability.
    Monoubiquitinated at Lys-21 and/or Lys-22 by UBE2D3. Ubiquitin chain elongation is then performed by CDC34 in cooperation with the SCF(FBXW11) E3 ligase complex, building ubiquitin chains from the UBE2D3-primed NFKBIA-linked ubiquitin. The resulting polyubiquitination leads to protein degradation. Also ubiquitinated by SCF(BTRC) following stimulus-dependent phosphorylation at Ser-32 and Ser-36.
    Deubiquitinated by porcine reproductive and respiratory syndrome virus Nsp2 protein, which thereby interferes with NFKBIA degradation and impairs subsequent NF-kappa-B activation.
  • 細胞内局在
    Cytoplasm. Nucleus. Shuttles between the nucleus and the cytoplasm by a nuclear localization signal (NLS) and a CRM1-dependent nuclear export.
  • Information by UniProt
  • 参照データベース
  • 別名
    • I kappa B alpha antibody
    • I-kappa-B-alpha antibody
    • IkappaBalpha antibody
    • IkB-alpha antibody
    • IKBA antibody
    • IKBA_HUMAN antibody
    • IKBalpha antibody
    • MAD 3 antibody
    • MAD3 antibody
    • Major histocompatibility complex enhancer-binding protein MAD3 antibody
    • NF kappa B inhibitor alpha antibody
    • NF-kappa-B inhibitor alpha antibody
    • NFKBI antibody
    • NFKBIA antibody
    • Nuclear factor of kappa light chain gene enhancer in B cells antibody
    • Nuclear factor of kappa light polypeptide gene enhancer in B cells inhibitor alpha antibody
    see all

Anti-IKB alpha antibody 画像



  • Predicted band size : 36 kDa
    Observed band size : 38 kDa (why is the actual band size different from the predicted?)

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: NFKBIA knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: Jurkat whole cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab95338 observed at 38 kDa. Red - loading control, ab18058, observed at 130 kDa.

    ab95338 was shown to specifically react with NFKBIA when NFKBIA knockout samples were used. Wild-type and NFKBIA knockout samples were subjected to SDS-PAGE. Ab95338 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • All lanes : Anti-IKB alpha antibody (ab95338) at 1 µg/ml

    Lane 1 : Human kidney tissue lysate - total protein (ab30203)
    Lane 2 : Human spleen tissue lysate - total protein (ab29699)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 36 kDa
    Observed band size : 36 kDa


    Exposure time : 4 minutes

  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 36 kDa


    Exposure time : 10 secondsAb95338 recognizes the tagged recombinant IKB alpha protein (ab59981) which has an expected molecular weight of 61kDa.
  • ICC/IF image of ab95338 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab95338, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

Anti-IKB alpha antibody (ab95338) 使用論文

ab95338 has not yet been referenced specifically in any publications.

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