antibody

I-kappa-Ba (Tyr-42), phospho-specific

MBS472028

0.1 ml

380 USD

polyclonal

rabbit

nonconjugated

phosphorylated

western blot, ELISA, immunoprecipitation

Antibody

I-kappa-Ba (Tyr-42), phospho-specific

I-kappa-Ba (Tyr-42)

NF-kappa-B inhibitor alpha; IKB-alpha; IkappaBalpha; Major histocompatibility complex enhancer-binding protein; MAD3; IkBa

Polyclonal

Rabbit

Human, Rat, Mouse

This antibody was cross-adsorbed to phospho-tyrosine coupled to agarose then affinity purified using phospho- I-kappa-B-beta (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 I-kappa-B-alpha.

Affinity Purified

Rabbit polyclonal, affinity-purified antibody is supplied in 100 ul phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide.

Store at -20 degree C. Do not aliquot. Stable for 1 year.

ELISA, Western Blot, Immunoprecipitation

ELISA: 1:2000. Immunoprecipitation: 5-10ml. Western Blot: 1:1000

Immunogen: IkBa (Tyr-42) synthetic peptide (coupled to KLH) corresponding to amino acid residues around tyrosine 42 of human IkBa. This peptide sequence has low homology to other IkB proteins, but does have some homology to unrelated proteins that may have a conserved tyrosine phosphorylation motif.

The NF-kB/Rel transcription factors are present in the cytosol in an inactive state complexed with the inhibitory IkB proteins. Activation of IkBa ?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-kB. This pathway of IkBa regulation occurs in response to various NF-kB-activating agents, such as TNF-alpha, interleukins, LPS, and irradiation. An alternative pathway for IkBa 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 IkB-alpha and NF-kB-activation. In contrast, Tyr-305 phosphorylation by c-Abl has been implicated in IkB-alpha nuclear translocation and inhibition of NF-kB-activation. Thus, tyrosine phosphorylation of IkB-alpha may be an important regulatory mechanism in NF-kB signaling.

Bui, N.T. et al. (2001) J Cell Biol 152(4):753. (Background). Waris et al. (2003) J Biol Chem 278(42):40778. (Background). Naidu, S. et al. (2008) J Immunol. 181:4113. (WB: Mouse RAW264.7). Sethi, G. et al. (2007) Oncogene 26(52):7324. (WB: H. epithelial H1299, Y42F). Boosani, C.S. et al. (2007) Blood 110(4):1168. (WB: M. lung endothelial, MLEC)

0.1 ml

380 USD