Product nameAnti-IKB alpha (phospho S32) antibody [EPR3148]
See all IKB alpha primary antibodies
DescriptionRabbit monoclonal [EPR3148] to IKB alpha (phospho S32)
Tested applicationsSuitable for: Dot blot, WB, IPmore details
Unsuitable for: Flow Cyt,ICC or IHC-P
Species reactivityReacts with: Human
Synthetic peptide within Human IKB alpha (phospho S32). The exact sequence is proprietary.
Database link: P25963
- WB: HeLa cell lysate, HeLa cell lysates treated with Calyculin A and TNF-a IP: HeLa treated with TNF-alpha whole cell lysate.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.
This product is a recombinant rabbit monoclonal antibody.
Storage instructionsShipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol, 0.05% BSA, 50% Tissue culture supernatant
PurityTissue culture supernatant
Our Abpromise guarantee covers the use of ab92700 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/500 - 1/10000. Predicted molecular weight: 36 kDa.|
|IP||1/10 - 1/100.|
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.
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 localizationCytoplasm. Nucleus. Shuttles between the nucleus and the cytoplasm by a nuclear localization signal (NLS) and a CRM1-dependent nuclear export.
- Information by UniProt
- I kappa B alpha antibody
- I-kappa-B-alpha antibody
- IkappaBalpha antibody
All lanes : Anti-IKB alpha (phospho S32) antibody [EPR3148] (ab92700) at 1/500 dilution
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with TNF-a at 20 ng/mL for 5 minutes. Whole cell lysates
Lane 3 : HeLa treated with TNF-a at 20 ng/mL for 5 minutes. Whole cell lysates. Then the membrane was incubated with phosphatase.
Lysates/proteins at 15 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 36 kDa
Observed band size: 36 kDa
Exposure time: 3 minutes
Blocking/Diluting buffer and concentration 5% NFDM/TBST
ab92700 at 1/20 dilution immunoprecipitating IKB alpha (phospho S32) in HeLa (human cervix adenocarcinoma) treated with 20ng/mL TNF-alpha for 60 minutes, whole cell lysate, observed at 36 kDa (lanes 1 and 2).
Lane 1 (input): HeLa treated with 20ng/mL TNF-alpha for 60 minutes, whole cell lysate, 10μg.
Lane 2 (+): ab92700 + HeLa treated with 20ng/mL TNF-alpha for 60 minutes, whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab92700 in HeLa treated with 20ng/mL TNF-alpha for 60 minutes, whole cell lysate.
For western blotting, ab92700 at 1/200 dilution followed by VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.
Blocking/Diluting buffer and concentration: 5% NFDM/TBST.
All lanes : Anti-IKB alpha (phospho S32) antibody [EPR3148] (ab92700) at 1/1000 dilution
Lane 1 : HeLa cell lysate untreated
Lane 2 : HeLa cell lysate treated with Calyculin A and TNF-a
Lysates/proteins at 10 µg per lane.
All lanes : HRP-labeled goat anti-rabbit at 1/2000 dilution
Predicted band size: 36 kDa
Dot blot analysis of IKB alpha (phospho S32) phospho peptide (Lane 1) and IKB alpha non-phospho peptide (Lane 2) labeling IKB alpha (phospho S32) with ab92700 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG) (H+L) at 1/10000 was used as the secondary antibody.
Blocking and diluting buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
This product has been referenced in:
- Yu T et al. Ligustrazine protects chondrocyte against IL-1ß induced injury by regulation of SOX9/NF-?B signaling pathway. J Cell Biochem 119:7419-7430 (2018). Read more (PubMed: 29806884) »
- Wei X et al. Hint1 expression inhibits proliferation and promotes radiosensitivity of human SGC7901 gastric cancer cells. Oncol Lett 16:2135-2142 (2018). Read more (PubMed: 30008911) »