Overview

  • Product nameAnti-IKB alpha antibody [6A920]
    See all IKB alpha primary antibodies
  • Description
    Mouse monoclonal [6A920] to IKB alpha
  • SpecificityAb12134 recognises IKB alpha.
  • Tested applicationsSuitable for: IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Hamster, Human, African Green Monkey
  • 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.

Properties

Applications

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
IHC-P Use at an assay dependent concentration.
WB Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 40 kDa (predicted molecular weight: 35.6 kDa).

Target

  • FunctionInhibits 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 diseaseEctodermal dysplasia, anhidrotic, with T-cell immunodeficiency autosomal dominant
  • Sequence similaritiesBelongs 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 localizationCytoplasm. Nucleus. Shuttles between the nucleus and the cytoplasm by a nuclear localization signal (NLS) and a CRM1-dependent nuclear export.
  • Information by UniProt
  • Database links
  • 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

Anti-IKB alpha antibody [6A920] images

  • All lanes : Anti-IKB alpha antibody [6A920] (ab12134) at 1/1000 dilution

    Lane 1 : Human B cell lysate treated with CD40L for 4 hours
    Lane 2 : Human B cell lysate treated with CD40L for 4 hours
    Lane 3 : Human B cell lysate treated with CD40L for 4 hours

    Lysates/proteins at 10 µg per lane.

    Secondary
    HRP-conjugated sheep anti-mouse IgG polyclonal at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 35.6 kDa
    Observed band size : 37 kDa (why is the actual band size different from the predicted?)


    Exposure time : 10 minutes

    This image is courtesy of an anonymous Abreview

    See Abreview

  • ab12134 at 1/250 staining human adrenal tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was blocked in serum before incubation with the primary antibody for 30 minutes at 22°C. An HRP conjugated goat anti-mouse antibody was used as the secondary.

    See Abreview



  • Predicted band size : 35.6 kDa

    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.

References for Anti-IKB alpha antibody [6A920] (ab12134)

This product has been referenced in:
  • 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 . Read more (PubMed: 23824821) »

See 1 Publication for this product

Product Wall

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
Username

Abcam user community

Verified customer

Submitted Oct 27 2014

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 ...

Read More
Abcam has not validated the combination of species/application used in this Abreview.
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
Username

Dr. Mahesh Shivananjappa

Verified customer

Submitted Jun 21 2012

Abcam has not validated the combination of species/application used in this Abreview.
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
Username

Dr. Mahesh Shivananjappa

Verified customer

Submitted Dec 08 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Dr. Mahesh Shivananjappa

Verified customer

Submitted Nov 14 2011

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
Username

Abcam user community

Verified customer

Submitted Oct 30 2007

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
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Abcam user community

Verified customer

Submitted Oct 30 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

Submitted Oct 09 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

Submitted Oct 05 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

Submitted Oct 05 2007

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"