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PhosphoSitePlus® Resource

  • Additional protein information
  • Analytical tools


Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb 2697 20 µl
Western Blotting Immunohistochemistry Flow Cytometry
H M R Mk 85 IKK-alpha 87 IKK-beta Rabbit IgG
IKKα (3G12) Mouse mAb 11930 20 µl
Western Blotting Immunofluorescence Flow Cytometry
H Mk 85 Mouse IgG1
Phospho-NF-κB2 p100 (Ser866/870) Antibody 4810 20 µl
Western Blotting Immunoprecipitation
H M 110 Rabbit 
NF-κB2 p100/p52 Antibody 4882 20 µl
Western Blotting Immunoprecipitation
H M R Mk 52 (mature). 120 (precursor). Rabbit 
NIK Antibody 4994 20 µl
Western Blotting
H M 125 Rabbit 
RelB (C1E4) Rabbit mAb 4922 20 µl
Western Blotting Immunoprecipitation
H M R Mk 70 Rabbit IgG
TRAF2 Antibody 4712 20 µl
Western Blotting
H M R Mk 53 Rabbit 
TRAF3 Antibody 4729 20 µl
Western Blotting
H M R Mk 62 Rabbit 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
Western Blotting
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl
Western Blotting

Product Description

This kit contains reagents to examine the activation state and total protein levels of key components in the noncanonical NF-κB pathway: TRAF2, TRAF3, NIK, IKKα, p100, and RelB.

Specificity / Sensitivity

The Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb detects IKKα only when phosphorylated at Ser176/180 and IKKβ only when phosphorylated at Ser177/181. The Phospho-NF-κB2 p100 (Ser866/870) Antibody detects transfected levels of NF-κB2 p100 only when phosphorylated at Ser866/870. The TRAF2, TRAF3, IKKα, RelB, and p100/p52 antibodies detect endogenous levels of total protein of their respective targets. The NIK antibody detects transfected levels of NIK regardless of modification state.

Source / Purification

Antibodies to phospho-IKKα/ß (Ser176/180) and phospho-p100 (Ser866/870) are produced by immunizing rabbits with synthetic phosphopeptides corresponding to amino acids surrounding the indicated residues of human IKKα and NFκB2 p100, respectively. Antibodies to TRAF2, TRAF3, NIK,, RelB, and p100/p52 are produced by immunizing rabbits with synthetic peptides corresponding to amino acids at the carboxy terminus of TRAF2, in a central region of TRAF3, adjacent to Gly659 of human NIK, surrounding Ser424 of human RelB, and at the amino terminus of human p100/p52, respectively. Monoclonal antibody to IKKα is produced by immunizing animals with a recombinant protein specific to a fragment of human IKKα protein. Polyclonal antibodies are purified by Protein A and peptide affinity chromatography.

Transcription factors of the nuclear factor κB (NF-κB)/Rel family play a pivotal role in inflammatory and immune responses (1,2). There are five family members in mammals: RelA, RelB, c-Rel, NF-κB1 (p105/p50) and NF-κB2 (p100/p52). Both p105 and p100 are proteolytically processed by the proteasome to produce p50 and p52, respectively. The p50 and p52 products form dimeric complexes with Rel proteins. While p50 associates with many of the NF-κB family members, p52 tends to form dimers primarily with RelB. A plethora of stimuli such TNFα and LPS induce the canonical NF-κB pathway, characterized by the activation of the classical IκB Kinase (IKK) complex (containing IKKα, IKKβ, IKKγ, and ELKS), which then phosphorylates inhibitory IκB molecules, targeting them for rapid degradation through a ubiquitin-proteasome pathway (3).

The noncanonical pathway, triggered by BAFF, CD40L, and certain other stimuli, is based on the inducible phosphorylation and proteasome-mediated partial degradation of NF-κB2 p100 to p52, a process regulated by the NF-κB Inducing Kinase (NIK) and IKKα, but not IKKβ or IKKγ (4-6). NIK phosphorylates IKKα at Ser176/180 (6) and p100 at Ser866/870, then recruits IKKα to p100 where IKKα phosphorylates additional residues in the N- and C-terminus (8), leading to the ubiquitination and processing of p100 (9). The TNF Receptor Associated Factor molecules TRAF2 and TRAF3 have been shown to be negative regulators of the noncanonical pathway (10, 11), and their differential binding to receptors may also play a role in determining whether transduced signals activate the canonical pathway, noncanonical pathway, or both (12). TRAF3 promotes the rapid turnover of NIK in resting cells, and its activation-induced degradation is a key regulatory point in the pathway (13). This pathway is required for B cell maturation and activation, proper architecture of peripheral lymphoid tissue, and safeguards against autoimmunity (14).

1.  Baeuerle, P.A. and Henkel, T. (1994) Annu Rev Immunol 12, 141-79.

2.  Baeuerle, P.A. and Baltimore, D. (1996) Cell 87, 13-20.

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

4.  Xiao, G. et al. (2001) EMBO J 20, 6805-15.

5.  Ling, L. et al. (1998) Proc Natl Acad Sci USA 95, 3792-7.

6.  Xiao, G. et al. (2004) J Biol Chem 279, 30099-105.

7.  Senftleben, U. et al. (2001) Science 293, 1495-9.

8.  Liang, C. et al. (2006) Cell Signal 18, 1309-17.

9.  Xia, Z.P. and Chen, Z.J. (2005) Sci STKE 2005, pe7.

10.  Xiao, G. et al. (2001) Mol Cell 7, 401-9.

11.  Liao, G. et al. (2004) J Biol Chem 279, 26243-50.

12.  Morrison, M.D. et al. (2005) J Biol Chem 280, 10018-24.

13.  Qing, G. et al. (2005) J Biol Chem 280, 40578-82.

14.  Xiao, G. et al. (2006) Cytokine Growth Factor Rev 17, 281-93.

Entrez-Gene Id 1147 , 4791 , 9020 , 5971 , 7186 , 7187
Swiss-Prot Acc. O15111 , Q00653 , Q99558 , Q01201 , Q12933 , Q13114

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. 7,429,487, foreign equivalents, and child patents deriving therefrom.

NF-κB Non-Canonical Pathway Antibody Sampler Kit