Synthetic peptide corresponding to Human IKB alpha (phospho Y305) conjugated to Keyhole Limpet Haemocyanin (KLH). Synthetic peptide conjugated to KLH corresponding to amino acid residues around tyrosine 305 of human IKB alpha. This peptide sequence has low homology to other IKB proteins.
A431 and Jurkat cells treated with pervanadate.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/500. Detects a band of approximately 38 kDa.
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 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.
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Anti-IKB alpha (phospho Y305) antibody images
Western blot - IKB alpha (phospho Y305) antibody (ab24784)
Western blot analysis of A431 cells treated with pervanadate (1 mM) for 30 minutes (30 ug/lane). Blots were probed with anti-IkappaBalpha (lane 1), anti-phospho-IkappaBalpha?(Tyr-42) (ab24783; lanes 2-5), or anti-phospho-IkappaBalpha (Tyr-305) (ab24784; lanes 6-9). Both anti-phospho-IkappaBalpha (Tyr-42) and anti-phospho-IkappaBalpha (Tyr-305) were used in the presence of no blocking peptide (lane 2 & 6), phospho-IkappaBalpha (Tyr-42) peptide (lane 3 & 8), phospho-IkappaBalpha (Tyr-305) peptide (lane 4 & 7), or BSA conjugated to phospho-tyrosine (lane 5 & 9). Peptides and BSA-pTyr were used at 1 ug/ml.
References for Anti-IKB alpha (phospho Y305) antibody (ab24784)
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