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Products:Signal Transduction >> Signaling Pathway >> Nuclear Signaling >> NFkB Pathway
Overview
Product name
Anti-IKB alpha (phospho S32 + S36) antibody
See all IKB alpha products (25) ...
Description
Rabbit polyclonal to IKB alpha (phospho S32 + S36)
Tested applications
Cross reactivity
Reacts with
Human
Predicted to work with
Mouse, Rat, Chicken, Pig
Immunogen
Synthetic phosphopeptide derived from the region of human IKBa that contains serines 32 and 36.
Positive control
Jurkat cells pretreated for 30 minutes with N-acetyl-leucinyl-leucinyl-norleucinal (ALLN) and stimulated with TNF alpha.
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: 0.05% Sodium Azide
Constituents: 50% Glycerol, PBS, 1mg/ml BSA. pH 7.3
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Purification notes
The antibody has been negatively pre-adsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated IKBa. The final product is generated by affinity chromatography using a IKBa-derived peptide that is phosphorylated at serines 32 and 36.
Clonality
Polyclonal
Isotype
IgG
-
Western blot - Anti-IKB alpha (phospho S32 + S36) antibody (ab5682)
(enlarge)
Applications
Show applications key
Our Abpromise guarantee covers the use of ab5682 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
WB: 1/1000Detects a band of approximately 40 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 tranlocate to the nucleus and activate transcription.
Involvement in disease
Defects in NFKBIA are the cause of ectodermal dysplasia anhidrotic with T-cell immunodeficiency autosomal dominant (ADEDAID) [MIM:612132]. Ectodermal dysplasia defines a heterogeneous group of disorders due to abnormal development of two or more ectodermal structures. ADEDAID is an ectodermal dysplasia associated with decreased production of pro-inflammatory cytokines and certain interferons, rendering patients susceptible to infection.
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.
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 polyubiquitinatin leads to protein degredation. 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 interefers with NFKBIA degradation and impairs subsequenbt 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 accessionP25963
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- I kappa B alpha antibody
- I(Kappa)B(alpha) antibody
- I-kappa-B-alpha antibody
see all
Database links
- Entrez Gene: 396093 Chicken
- Entrez Gene: 4792 Human
- Entrez Gene: 18035 Mouse
- Entrez Gene: 406188 Pig
- Entrez Gene: 25493 Rat
- Omim: 164008 Human
- SwissProt: Q91974 Chicken
- SwissProt: P25963 Human
see all
Anti-IKB alpha (phospho S32 + S36) antibody images:
Western blot - Anti-IKB alpha (phospho S32 + S36) antibody (ab5682)
Peptide Competition and Induction: Extracts prepared from ALLN pretreated Jurkat cells left unstimulated (1, 2) or stimulated with TNF alpha (3 7) were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer for one hour at room temperature and incubated with anti-IKBa pan (1, 3) or ab5682 antibody (2, 4-7) overnight at 4°C in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1-4), the non phosphopeptide corresponding to the immunogen (5), a generic phosphoserine-containing peptide (6), or, the phosphopeptide immunogen (7). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG HRP conjugate and bands were detected using the Pierce SuperSignal method. The data show that the phosphorylation of IKBa is induced by the addition of TNF alpha and that the peptide corresponding to IKBa [pSpS32/36] blocks the antibody signal, thereby demonstrating the specificity of the antibody. The data also show that treatment with TNF alpha leads to degradation of IKBa protein (compare lanes 1 and 3), as expected, even in the presence of ALLN.
References for Anti-IKB alpha (phospho S32 + S36) antibody (ab5682)
This product has been referenced in:
- Numaguchi Yet al. Ablation of angiotensin IV receptor attenuates hypofibrinolysis via PAI-1 downregulation and reduces occlusive arterial thrombosis. Arterioscler Thromb Vasc Biol 29:2102-8 (2009).Read more (PubMed: 19745198) »
See 1 publication for this product
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"