Catalog # IK6320

IκBα Phospho-Regulation Antibody Kit

Antibody Sampler Kit

DATASHEET  




$395

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.

References
Finco, T.S. et al. (1994) Proc. Natl. Acad. Sci. USA 91:11884.
Bui, N.T. et al. (2001) J Cell Biol 152(4):753.
Waris et al. (2003) J Biol Chem 278(42):40778.

Western blot analysis of Jurkat cells untreated (lanes 1 & 3) or treated with TNFα (1 nM). The blots were probed with anti-IκBα (lanes 1 & 2) or anti-IκBα (Ser-32/Ser-36) (lanes 3 & 4).

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).



Mouse monoclonal and rabbit polyclonal antibodies are supplied in phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. The secondary reagents are supplied in the same buffer without azide. Store all at –20°C. Stable for 1 year.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.



*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

Product References:

IP1031 Cullen, S. et al. (2015) Biomolecules. 5(1):95 WB: ILU-18 cells
IP1041 Cullen, S. et al. (2015) Biomolecules. 5(1):95 WB: ILU-18 cells
IP1861 Karki, R. et al. (2014) Free Radic Biol Med. 71:256 WB: HEK-Blue mTLR4
RS3251 Kawasaki, H. et al. (2013) World J Gastroenter. 19(17):2629. WB, ICC: mouse intestinal myofibroblasts
MS3001 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot
RS3251 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot
IP1031 Wright, C.J. et al (2009) AJP Lung Cell Mol Phy 296(3):L296. WB: rat fibroblast
IP1031 Naidu, S. et al. (2008) J Immunol. 181:4113. WB: mouse RAW264.7
IP1031 Boosani, C.S. et al. (2007) Blood 110(4):1168. WB: mouse lung endothelial, MLEC
IP1031 Sethi, G. et al. (2007) Oncogene 26(52):7324. WB: human epithelial H1299, Y42F

This kit contains:

CATALOG#DESCRIPTIONSIZEAPPLICATIONSSPECIES REACTIVITYMW (kDa)
IκBα (C-terminus) Rabbit pAb50 μlWB, EHu, Rt, Ms38
IκBα (Ser-32/Ser-36), phospho-specific Mouse mAb50 μlWB, E, IPHu, Rt, Ms38
IκBα (Tyr-42) Rabbit pAb 50 μlWB, E, IPHu, Rt, Ms38
IκBα (Tyr-305), phospho-specific Rabbit pAb50 μlWB, EHu, Rt, Ms38
Anti-Mouse Ig:HRP Donkey pAb 100 μlWB, EMs
Anti-Rabbit Ig Light ChainSpecific:HRP Mouse mAb 100 μlWB, E, ICC, IHCRb
KIT SUMMARY

The IκBα phospho-regulation antibody sampler kit can be used to detect IκBα phosphorylation on Ser-32/Ser-36, Tyr-42, and Tyr-305. The kit also includes a polyclonal antibody to monitor total IκBα expression levels and secondary reagents for detection of these antibodies.