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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 [6A920] (ab12134)

  • Datasheet
  • SDS
Reviews (14)Q&A (1)References (3)

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Western blot - Anti-IKB alpha antibody [6A920] (ab12134)
  • Western blot - Anti-IKB alpha antibody [6A920] (ab12134)

Key features and details

  • Mouse monoclonal [6A920] to IKB alpha
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG1

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Protein
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Overview

  • Product name

    Anti-IKB alpha antibody [6A920]
    See all IKB alpha primary antibodies
  • Description

    Mouse monoclonal [6A920] to IKB alpha
  • Host species

    Mouse
  • Specificity

    Ab12134 recognises IKB alpha.
  • Tested applications

    Suitable for: WBmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    6xHis-tagged fusion protein, corresponding to amino acids 105-291 of Human IKB alpha.

  • Positive control

    • Jurkat + PMA (10 nm/ml) + ionomycin (1 µM) cell extract.
  • 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. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer

    Preservative: 0.02% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Purification notes

    Affinity purified.
  • Clonality

    Monoclonal
  • Clone number

    6A920
  • Isotype

    IgG1
  • 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-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
  • Recombinant Protein

    • Recombinant Human IKB alpha protein (ab113133)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab12134 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 (8)
Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 40 kDa (predicted molecular weight: 35.6 kDa).
Notes
WB
Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 40 kDa (predicted molecular weight: 35.6 kDa).

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: 4792 Human
    • Omim: 164008 Human
    • SwissProt: P25963 Human
    • Unigene: 81328 Human
    • 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 [6A920] (ab12134)
      Western blot - Anti-IKB alpha antibody [6A920] (ab12134)

      Lane 1: Wild type HAP1 whole cell lysate (20 µg)
      Lane 2: IKB alpha knockout HAP1 whole cell lysate (20 µg)
      Lane 3: HeLa whole cell lysate (20 µg)
      Lane 4: HepG2 whole cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab12134 observed at 38 kDa. Red - loading control, ab176560, observed at 50 kDa.

      ab12134 was shown to specifically react with IKB alpha in wild type cells as signal was lost in IKB alpha knockout cells. Wild-type and IKB alpha knockout samples were subjected to SDS-PAGE. Ab12134 and ab176560 (Rabbit anti alpha Tubulin loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-IKB alpha antibody [6A920] (ab12134)
      Western blot - Anti-IKB alpha antibody [6A920] (ab12134)

      ab12134 detects both phosphorylated and non-phosphorylated forms of IKB alpha by Western blot. Jurkat cells (1E7) were treated for indicated time periods with 10 nm/ml PMA, 1 µM ionomycin and ab12134 (1/10000 dilution). 10 µl of cytoplasmic (C) and nuclear extract (N) were resolved on a 8.75% SDS-PAGE and transferred to Immobilon membrane.

      ab12134 detects both phosphorylated and non-phosphorylated forms of IKB alpha by Western blot. Jurkat cells (1E7) were treated for indicated time periods with 10 nm/ml PMA, 1 µM ionomycin and ab12134 (1/10000 dilution). 10 µl of cytoplasmic (C) and nuclear extract (N) were resolved on a 8.75% SDS-PAGE and transferred to Immobilon membrane.

    Protocols

    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (3)

    Publishing research using ab12134? Please let us know so that we can cite the reference in this datasheet.

    ab12134 has been referenced in 3 publications.

    • Yang K  et al. Hirudin Ameliorates Renal Interstitial Fibrosis via Regulating TGF-ß1/Smad and NF-?B Signaling in UUO Rat Model. Evid Based Complement Alternat Med 2020:7291075 (2020). PubMed: 32714415
    • Ming Y  et al. Molecular hydrogen improves type 2 diabetes through inhibiting oxidative stress. Exp Ther Med 20:359-366 (2020). PubMed: 32537002
    • Chang MS  et al. Epstein-Barr virus-encoded BARF1 promotes proliferation of gastric carcinoma cells through regulation of NF-?B. J Virol 87:10515-23 (2013). WB ; Human . PubMed: 23824821

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 15 Abreviews or Q&A

    Western blot abreview for Anti-IKB alpha antibody [6A920]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    10 µg
    Gel Running Conditions
    Reduced Denaturing (10)
    Sample
    Human Cell lysate - whole cell (B cells)
    Specification
    B cells
    Treatment
    CD40L for 4 hours
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 27 2014

    Question

    ReviewID: xxx AbID: 12134, Anti-IKB alpha antibody [6A920] Rating: Below Average Image: Anti-IKB alpha antibody [6A920] for Immunocytochemistry/ Immunofluorescence (Mouse) Sample: Species: Mouse Type: Cultured Cells Specification: RAW 264.7 Application: Application: Fixative: Paraformaldehyde Permeabilization: Yes Blocking step: BSA as blocking agent for 1 hour at 4°C Other detail: Dilution: 1/100 Incubation time: 16 hours at 4°C Diluent: 2% BSA in PBS Secondary Antibody: Name: Non-Abcam Antibody was used: Alexa Flour488 Goat Anti Mouse IgG (H+L) Conjugation: Alexa Fluor® 488 Dilution: 1/500 Additional Data:

    Read More

    Abcam community

    Verified customer

    Asked on Jun 21 2012

    Answer

    Thank you for submitting an Abreview for ab12134. As you may have noticed, your review has now been published on our website.
    Since you obtained poor results using the antibody in a untested species/application, we would like to follow up on this to see if we can possibly improve the results you are seeing with this antibody.
    Based on the information in your review, I feel that altering the following step in your protocol may help improve your results:
    - try a different fixation reagent (e.g. acetone)
    - increase the permeabilization time
    - use more primary antibody (e.g. 1/50)
    If you decide to follow these suggestions, please let me know if they are helpful.
    I wish you good luck with your research.

    Read More

    Abcam Scientific Support

    Answered on Jun 21 2012

    Immunocytochemistry/ Immunofluorescence abreview for Anti-IKB alpha antibody [6A920]

    Below Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cultured Cells (RAW 264.7)
    Specification
    RAW 264.7
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.1% Triton-X100 in 2% BSA for 15min
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 4°C
    Read More

    Dr. Mahesh Shivananjappa

    Verified customer

    Submitted Jun 21 2012

    Flow Cytometry abreview for Anti-IKB alpha antibody [6A920]

    Below Average
    Abreviews
    Abreviews
    abreview image
    Application
    Flow Cytometry
    Sample
    Human Cell (Monocytes from Apheresis)
    Specification
    Monocytes from Apheresis
    Fixation
    BD Cytofix/Cytoperm 20 min
    Permeabilization
    Yes - BD Cytofix/Cytoperm 20 min
    Gating Strategy
    Monocytes
    Read More

    Dr. Mahesh Shivananjappa

    Verified customer

    Submitted Dec 08 2011

    Western blot abreview for Anti-IKB alpha antibody [6A920]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Platelets)
    Loading amount
    20 µg
    Specification
    Platelets
    Treatment
    A23187 for 1 h
    Gel Running Conditions
    Reduced Denaturing (4-20%)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More

    Dr. Mahesh Shivananjappa

    Verified customer

    Submitted Nov 14 2011

    Immunoprecipitation abreview for Anti-IKB alpha antibody [6A920]

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunoprecipitation
    Sample
    Human Cell lysate - whole cell (U2OS cell line)
    Total protein in input
    500 µg
    Specification
    U2OS cell line
    Immuno-precipitation step
    Protein G
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 30 2007

    Immunoprecipitation abreview for Anti-IKB alpha antibody [6A920]

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunoprecipitation
    Sample
    Mouse Cell lysate - whole cell (3T3 cell line)
    Total protein in input
    500 µg
    Specification
    3T3 cell line
    Immuno-precipitation step
    Protein G
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 30 2007

    Western blot abreview for Anti-IKB alpha antibody [6A920]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Cell lysate - whole cell (PC12)
    Loading amount
    20 µg
    Specification
    PC12
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 09 2007

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-IKB alpha antibody [6A920]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (adrenal)
    Specification
    adrenal
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0
    Permeabilization
    No
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 22°C
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 05 2007

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-IKB alpha antibody [6A920]

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (adrenal)
    Specification
    adrenal
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0
    Permeabilization
    No
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 22°C
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 05 2007

    1-10 of 15 Abreviews or Q&A

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