Our IκBα (Tyr-42) rabbit polyclonal phosphospecific primary antibody from PhosphoSolutions is produced in-house. It detects human, mouse, and rat IκBα (Tyr-42) and is antigen affinity purified. It is great for use in WB, IP.
Western blot analysis of A431 cells treated with pervanadate (1 mM) for 30 min. Blots were probed with anti-IκBα (lane 1), anti-IκBα (Tyr-42) (IP1031; lanes 2-5), or anti-IκBα (Tyr-305) (IP1041; lanes 6-9). In some lanes, the antibodies were used in the absence (lane 2 & 6) or presence of IκBα (Tyr-42) (lane 3 & 8) or IκBα (Tyr-305) (lane 4 & 7) blocking peptides, or BSA conjugated to phospho-tyrosine (lane 5 & 9).
The NF-κB/Rel transcription factors are present in the cytosol in an inactive state complexed with the inhibitory IκB proteins. Activation of IκBα occurs through both serine and tyrosine phosphorylation events. Activation through phosphorylation at Ser-32 and Ser-36 is followed by proteasome-mediated degradation, resulting in the release and nuclear translocation of active NF-κB. This pathway of IκBα regulation occurs in response to various NF-κB-activating agents, such as TNFα, interleukins, LPS, and irradiation. An alternative pathway for IκBα regulation occurs through tyrosine phosphorylation of Tyr-42 and Tyr-305. Tyr-42 is phosphorylated in response to oxidative stress and growth factors. This phosphorylation can lead to degradation of IκBα and NF-κB-activation. In contrast, Tyr-305 phosphorylation by c-Abl has been implicated in IκBα nuclear translocation and inhibition of NF-κB-activation. Thus, tyrosine phosphorylation of IκBα may be an important regulatory mechanism in NF-κB signaling.
Antigen Affinity Purified
Polyclonal
IgG
IP, WB
Rabbit
NFKBIA
38
IκBα (Tyr-42) synthetic peptide (coupled to KLH) corresponding to amino acid residues around tyrosine 42 of human IκBα. This peptide sequence has low homology to other IκB proteins, but does have some homology to unrelated proteins that may have a conserved tyrosine phosphorylation motif.
Human, Mouse, Rat
Storage at -20°C is recommended, as aliquots may be taken without freeze/thawing due to presence of 50% glycerol. Stable for at least 1 year at -20°C.
Liquid
PBS + 1 mg/ml BSA, 0.05% NaN3 and 50% glycerol
WB: 1:1000
IP: 1:100
Unconjugated
This antibody was cross-adsorbed to phospho-tyrosine coupled to agarose then affinity purified using phospho-IκBα (Tyr-42) peptide (without carrier). The antibody detects a 38 kDa* protein on SDS-PAGE immunoblots of A431 and Jurkat cells treated with pervanadate, but not in control cells. Due to homologies to tyrosine sites on other proteins, it is recommended that the antibody be used to detect phosphorylation of immunoprecipitated IκBα.
Phosphorylated
Tyr-42
Western blots performed on each lot.
For research use only. Not intended for therapeutic or diagnostic use. Use of all products is subject to our terms and conditions, which can be viewed on our website.
United States
After date of receipt, stable for at least 1 year at -20°C.
Zhao, X, et al. 2018. A role for the non-receptor tyrosine kinase ACK1 in TNF-alpha-mediated apoptosis and proliferation in human intestinal epithelial caco-2 cells. Cell biology international, 1097-1105.
Cullen, S., et al. 2015. Redox-regulated pathway of tyrosine phosphorylation underlies NF-κB induction by an atypical pathway independent of the 26S proteasome. Biomolecules, 95-112.
Zhang, Y, et al. 2018. Exogenous oxidants activate nuclear factor kappa B through Toll-like receptor 4 stimulation to maintain inflammatory phenotype in macrophage. Biochemical pharmacology, 104-118.
Wright, C.J., et al. 2009. Hyperoxia-induced NF-kappaB activation occurs via a maturationally sensitive atypical pathway. American Journal of Physiology. Lung Cellular and Molecular Physiology, L296-L306.
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