Anti-IKB alpha antibody [6A920] (ab12134)
- Datasheet
- References (1)
- Protocols
Overview
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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: 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.
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Positive control
- Jurkat + PMA (10 nm/ml) + ionomycin (1 µM) cell extract.
Properties
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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
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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
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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
modificationsPhosphorylated; 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. - Information by UniProt
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Database links
- Entrez Gene: 4792 Human
- Entrez Gene: 18035 Mouse
- Entrez Gene: 25493 Rat
- Omim: 164008 Human
- SwissProt: P25963 Human
- SwissProt: Q9Z1E3 Mouse
- SwissProt: Q63746 Rat
- Unigene: 81328 Human
see all -
Alternative names
- I kappa B alpha antibody
- I-kappa-B-alpha antibody
- IkappaBalpha antibody
see all
Images
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![Western blot - Anti-IKB alpha antibody [6A920] (ab12134)](https://www.abcam.com/ps/products/12/ab12134/Images/ab12134-290270-anti-ikb-alpha-antibody-6a920-western-blot.jpg)
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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKB alpha antibody [6A920] (ab12134)](https://www.abcam.com/ps/products/12/ab12134/Images/ab12134-19515-anti-ikb-alpha-antibody-6a920-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IKB alpha antibody [6A920] (ab12134)This image is courtesy of an anonymous Abreviewab12134 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.
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![Western blot - Anti-IKB alpha antibody [6A920] (ab12134)](https://www.abcam.com/ps/products/12/ab12134/Images/a4.jpg)
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
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.µ µ
Datasheets and documents
References
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) »
