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Signal Transduction Signaling Pathway Nuclear Signaling NFkB Pathway
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Validated using a knockout cell line

Anti-IKB alpha antibody (ab95338)

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Western blot - Anti-IKB alpha antibody (ab95338)
  • Western blot - Anti-IKB alpha antibody (ab95338)
  • Western blot - Anti-IKB alpha antibody (ab95338)
  • Immunocytochemistry/ Immunofluorescence - Anti-IKB alpha antibody (ab95338)

Key features and details

  • Rabbit polyclonal to IKB alpha
  • Suitable for: WB, ICC/IF
  • Knockout validated
  • Reacts with: Human, Recombinant fragment
  • Isotype: IgG

You may also be interested in

Protein
Product image
Recombinant Human IKB alpha protein (ab113133)
Secondary
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)

View more associated products

Overview

  • Product name

    Anti-IKB alpha antibody
    See all IKB alpha primary antibodies
  • Description

    Rabbit polyclonal to IKB alpha
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Human, Recombinant fragment
    Predicted to work with: Mouse, Rat, Sheep, Cow, Pig
  • Immunogen

    Synthetic peptide corresponding to Human IKB alpha aa 1-100 conjugated to keyhole limpet haemocyanin.
    Database link: P25963
    (Peptide available as ab105485)

  • Positive control

    • Recombinant Human IKB alpha protein (ab113133) can be used as a positive control in WB. 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.
  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • NFkB Pathway
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • NFkB pathway
    • Cancer
    • Signal transduction
    • Nuclear signaling
    • NFkB pathway
    • Immunology
    • Innate Immunity
    • TLR Signaling

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Positive Controls

    • Recombinant Human IKB alpha protein (ab113133)
  • Recombinant Protein

    • Recombinant Human IKB alpha protein (ab113133)

Applications

The Abpromise guarantee

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.

Application Abreviews Notes
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.
Notes
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.

Target

  • Function

    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.
  • Involvement in disease

    Ectodermal dysplasia, anhidrotic, with T-cell immunodeficiency autosomal dominant
  • Sequence similarities

    Belongs to the NF-kappa-B inhibitor family.
    Contains 5 ANK repeats.
  • Post-translational
    modifications

    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.
  • Cellular localization

    Cytoplasm. Nucleus. Shuttles between the nucleus and the cytoplasm by a nuclear localization signal (NLS) and a CRM1-dependent nuclear export.
  • Target information above from: UniProt accession P25963 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 282291 Cow
    • Entrez Gene: 4792 Human
    • Entrez Gene: 18035 Mouse
    • Entrez Gene: 406188 Pig
    • Entrez Gene: 25493 Rat
    • Omim: 164008 Human
    • SwissProt: P25963 Human
    • SwissProt: Q9Z1E3 Mouse
    • SwissProt: Q08353 Pig
    • SwissProt: Q63746 Rat
    • Unigene: 81328 Human
    • Unigene: 170515 Mouse
    • Unigene: 12550 Rat
    see all
  • Alternative names

    • 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

Images

  • Western blot - Anti-IKB alpha antibody (ab95338)
    Western blot - Anti-IKB alpha antibody (ab95338)

    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 in wild-type HAP1 cells. No band was observed when NFKBIA knockout samples were examined. 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 1/1000 dilution and 1/10,000 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 1/10,000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-IKB alpha antibody (ab95338)
    Western blot - Anti-IKB alpha antibody (ab95338)
    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
    All lanes : 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
  • Western blot - Anti-IKB alpha antibody (ab95338)
    Western blot - Anti-IKB alpha antibody (ab95338)
    Ab95338 recognizes the tagged recombinant IKB alpha protein (ab59981) which has an expected molecular weight of 61kDa.
  • Immunocytochemistry/ Immunofluorescence - Anti-IKB alpha antibody (ab95338)
    Immunocytochemistry/ Immunofluorescence - Anti-IKB alpha antibody (ab95338)
    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.

Protocols

  • Western blot protocols
  • Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (0)

Publishing research using ab95338? Please let us know so that we can cite the reference in this datasheet.

ab95338 has not yet been referenced specifically in any publications.

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