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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
IKKα (3G12) Mouse mAb 11930 40 µl
Western Blotting Immunofluorescence Flow Cytometry
H Mk 85 Mouse IgG1
IKKβ (D30C6) Rabbit mAb 8943 40 µl
Western Blotting Immunoprecipitation
H M R Mk 87 Rabbit IgG
Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb 2697 40 µl
Western Blotting Immunohistochemistry Flow Cytometry
H M R Mk 85 IKK-alpha 87 IKK-beta Rabbit IgG
Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb 3033 40 µl
Western Blotting Immunoprecipitation Immunofluorescence Flow Cytometry
H M R Hm Mk Pg 65 Rabbit IgG
IκBα (L35A5) Mouse mAb (Amino-terminal Antigen) 4814 40 µl
Western Blotting Immunoprecipitation Immunohistochemistry Immunofluorescence Flow Cytometry
H M R Mk B Pg GP 39 Mouse IgG1
Phospho-IκBα (Ser32) (14D4) Rabbit mAb 2859 40 µl
Western Blotting Immunoprecipitation
H M R Mk 40 Rabbit IgG
NF-κB p65 (D14E12) XP® Rabbit mAb 8242 40 µl
Western Blotting Immunoprecipitation Immunohistochemistry Immunofluorescence Flow Cytometry Chromatin Immunoprecipitation
H M R Hm Mk Dg 65 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
Western Blotting
Goat 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl
Western Blotting
All Horse 

Product Description

The NF-κB Pathway Sampler Kit contains reagents to examine the activation state and total protein levels of key proteins in the NF-κB pathway: IKKα, IKKβ, NF-κB p65/RelA and IκBα. The kit contains enough primary and secondary antibodies to perform four Western blot experiments per primary antibody.


Specificity / Sensitivity

Phospho-IKKα/β, phospho-NF-κB p65, and phospho-IκBα antibodies recognize endogenous levels IKKα/β, p65, and IκB-α, respectively, only when phosphorylated at the indicated residues. They do not cross-react with other family members at physiological levels. Total IKKα, IKKβ, p65 and IκBα antibodies recognize endogenous levels of their respective targets regardless of phosphorylation state and do not cross-react with other family members at physiological levels.


Source / Purification

Monoclonal antibodies are produced by immunizing animals with recombinant human proteins or synthetic peptides.

The transcriptional nuclear factor κB (NF-κB)/Rel transcription factors are present in the cytosol in an inactive state, complexed with the inhibitory IκB proteins. Activation occurs via phosphorylation of IκBα at Ser32 and Ser36, resulting in the ubiquitin-mediated proteasome-dependent degradation of IκBα and the release and nuclear translocation of active NF-κB dimers. The regulation of IκBβ and IκBε is similar to that of IκBα, however, the phosphorylation and degradation of these proteins occurs with much slower kinetics. Phosphorylation of IκBβ occurs at Ser/Thr19 and Ser23, while IκBε can be phosphorylated at Ser18 and Ser22. The key regulatory step in this pathway involves activation of a high molecular weight IkappaB kinase (IKK) complex, consisting of three tightly associated IKK subunits. IKKα and IKKβ serve as the catalytic subunits of the kinase. Activation of IKK depends on phosphorylation at Ser177 and Ser181 in the activation loop of IKKβ (176 and 180 in IKKα). NF-κB-inducing kinase (NIK), TANK-binding kinase 1 (TBK1), and its homolog IKKε (IKKi), phosphorylate and activate IKKα and IKKβ.


The NF-κB family of transcription factors is comprised of five proteins in mammals, p65/RelA, c-Rel, RelB, NF-κB1 (p105/p50) and NF-κB2 (p100/p52). p105 and p100 are proteolytically processed to produce p50 and p52, respectively. The 50 kDa active form is produced through proteolytic processing following IKK-mediated phosphorylation of p105 at multiple sites (Ser922, 924, 928 and 933), while p100's processing to p52 is induced by phosphorylation of Ser864 and Ser868. The p50 and p52 products form dimeric complexes with Rel proteins, which are then able to bind DNA and regulate transcription. Phosphorylation of p65/RelA at Ser276 by PKA C and MSK1 enhances transcriptional activity. p65 phosphorylation at Ser536 regulates activation, nuclear localization, protein-protein interactions, and transcriptional activity. PMA-induced NF-κB transcriptional activity is dependent on the region of p65 containing the potential phosphorylation sites Ser457, Thr458, Thr464 and Ser468. Phosphorylation of Ser468 by GSK-3β inhibits basal p65 activity.


1.  Yamamoto, Y. and Gaynor, R.B. (2004) Trends Biochem. Sci. 29, 72-79.

2.  Ghosh, S. and Karin, M. (2002) Cell 109, S81-S96.

3.  Viatour, P. et al. (2005) Trends Biochem. Sci. 30, 43-52.


Entrez-Gene Id 4792 , 1147 , 3551 , 5970
Swiss-Prot Acc. P25963 , O15111 , O14920 , Q04206


For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 5,675,063.

9936
NF-κB Pathway Sampler Kit