RecombinantRabMAb

Recombinant Anti-IKB alpha (phospho S32) antibody [EPR3148] (ab92700)

Submit a review Q&A (1)References (35)
Western blot - Anti-IKB alpha (phospho S32) antibody [EPR3148] (ab92700)
  • Western blot - Anti-IKB alpha (phospho S32) antibody [EPR3148] (ab92700)
  • Immunoprecipitation - Anti-IKB alpha (phospho S32) antibody [EPR3148] (ab92700)
  • Western blot - Anti-IKB alpha (phospho S32) antibody [EPR3148] (ab92700)
  • Dot Blot - Anti-IKB alpha (phospho S32) antibody [EPR3148] (ab92700)
  • Anti-IKB alpha (phospho S32) antibody [EPR3148] (ab92700)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3148] to IKB alpha (phospho S32)
  • Suitable for: Dot blot, WB, IP
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-IKB alpha (phospho S32) antibody [EPR3148]
    See all IKB alpha primary antibodies

  • Description

    Rabbit monoclonal [EPR3148] to IKB alpha (phospho S32)

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Dot blot, WB, IPmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide within Human IKB alpha aa 1-100 (phospho S32). The exact sequence is proprietary.
    Database link: P25963

  • Positive control

    • WB: TNF-a treated HeLa and TNF-a treated MCF7 whole cell lysates; IP: TNF-a treated HeLa whole cell lysate.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Properties

Applications

The Abpromise guarantee

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.

Application Abreviews Notes
Dot blot
1/1000.
WB
1/1000. Predicted molecular weight: 36 kDa.

For unpurified use at 1/500 - 1/10,000
IP
1/20.

For unpurified use at 1/10 - 1/100
Notes
Dot blot
1/1000.
WB
1/1000. Predicted molecular weight: 36 kDa.

For unpurified use at 1/500 - 1/10,000
IP
1/20.

For unpurified use at 1/10 - 1/100

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.
  • 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

    Images

    • Western blot - Anti-IKB alpha (phospho S32) antibody [EPR3148] (ab92700)
      Western blot - Anti-IKB alpha (phospho S32) antibody [EPR3148] (ab92700)
      All lanes : Anti-IKB alpha (phospho S32) antibody [EPR3148] (ab92700) at 1/1000 dilution (Purified)

      Lane 1 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate
      Lane 2 : MCF7 (Human breast adenocarcinoma epithelial cell) treated with 20 ng/mL TNF-alpha for 8 hours whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 36 kDa
      Observed band size: 36 kDa



      Blocking/Diluting buffer: 5% NFDM/TBST

    • Western blot - Anti-IKB alpha (phospho S32) antibody [EPR3148] (ab92700)
      Western blot - Anti-IKB alpha (phospho S32) antibody [EPR3148] (ab92700)
      All lanes : Anti-IKB alpha (phospho S32) antibody [EPR3148] (ab92700) at 1/1000 dilution (Purified)

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
      Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with 20 ng/mL TNF-alpha for 5 minutes whole cell lysate

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 36 kDa
      Observed band size: 36 kDa



      Blocking/Diluting buffer: 5% NFDM/TBST

    • Immunoprecipitation - Anti-IKB alpha (phospho S32) antibody [EPR3148] (ab92700)
      Immunoprecipitation - Anti-IKB alpha (phospho S32) antibody [EPR3148] (ab92700)

      Purified ab92700 at 1:20 dilution (1μg) immunoprecipitating IKB alpha in HeLa treated with 20ng/mL TNF-alpha for 60 minutes whole cell lysate.

      Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) 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.

      Blocking Buffer and concentration: 5% NFDM/TBST.
      Diluting buffer and concentration: 5% NFDM/TBST.
      Observed band size: 36 kDa

    • Western blot - Anti-IKB alpha (phospho S32) antibody [EPR3148] (ab92700)
      Western blot - Anti-IKB alpha (phospho S32) antibody [EPR3148] (ab92700)
      All lanes : Anti-IKB alpha (phospho S32) antibody [EPR3148] (ab92700) at 1/500 dilution (unpurified)

      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.

      Secondary
      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

    • Dot Blot - Anti-IKB alpha (phospho S32) antibody [EPR3148] (ab92700)
      Dot Blot - Anti-IKB alpha (phospho S32) antibody [EPR3148] (ab92700)

      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 unpurified 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.

    • Anti-IKB alpha (phospho S32) antibody [EPR3148] (ab92700)
      Anti-IKB alpha (phospho S32) antibody [EPR3148] (ab92700)

    References (35)

    Publishing research using ab92700? Please let us know so that we can cite the reference in this datasheet.

    ab92700 has been referenced in 35 publications.

    • Ji C  et al. Propofol alleviates inflammation and apoptosis in HCY-induced HUVECs by inhibiting endoplasmic reticulum stress. Mol Med Rep 23:N/A (2021). PubMed: 33760174
    • Zhang X  et al. Exploring the five different genes associated with PKCa in bladder cancer based on gene expression microarray. J Cell Mol Med 25:1759-1770 (2021). PubMed: 33452764
    • Wang SH  et al. Insulin-like growth factor binding protein 3 promotes radiosensitivity of oral squamous cell carcinoma cells via positive feedback on NF-?B/IL-6/ROS signaling. J Exp Clin Cancer Res 40:95 (2021). PubMed: 33712045
    • Chen HL  et al. Galectin-7 downregulation in lesional keratinocytes contributes to enhanced IL-17A signaling and skin pathology in psoriasis. J Clin Invest 131:N/A (2021). PubMed: 33055419
    • Deji QZ  et al. Cross-talk between microRNA-let7c and transforming growth factor-ß2 during epithelial-to-mesenchymal transition of retinal pigment epithelial cells. Int J Ophthalmol 13:693-700 (2020). PubMed: 32420214
    View all Publications for this product

    Customer reviews and Q&As

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

    Question

    Thank you for your response. Sorry I realised I will be using the antibody in a WB after all. I am interested in using a Phospho antibody, was wondering if there is any difference / advantage in using either the Ser32 or the Ser32/36 antibody (eg ab5682 vs ab92700) ?

    Thanks for your help

    Read More
    Answer

    Thank you for your reply.

    Both mentioned products would detect the IKB alpha in WB and are guaranteed for human samples. If you want to use other samples than that, you can see on the data sheet, that the ab5682 is predicted to work with Mouse, Rat, Chicken, Pig. That means it has not been tested yet but if you like to try this antibody with one of these samples you would be eligible for the Abtrial. You can read more about our Abtrial programme https://www.abcam.com/index.html?pageconfig=resource&rid=11998&source=pagetrap&viapagetrap=abtrial.

    The ab92700 does not react with mouse and rat and we would not recommend to try this product with this kind of samples.

    I would also recommend to have a look at the specific references for each of the products cited on our webpage, as well as the Abreview for the ab5682. I hope this information will help you to decide, which of these antibodies might be best for you.

    If you have further questions, please do not hesitate to contact me again.

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com