Product Pathways - NF-kappaB Signaling

NF-κB Non-Canonical Pathway Antibody Sampler Kit #4888

• pathway
• more info
• application references
• datasheet PDF
• MSDS PDF
• protocols

Kit Includes	Quantity	Applications	Reactivity	MW (kDa)	Source
Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb # 2697	40 microliters	W IHC-P IHC-F	H M R Mk (B)	85 IKK-alpha 87 IKK-beta	Rabbit
IKKα Antibody # 2682	40 microliters	W IP	H M R Mk (B)	85	Rabbit
Phospho-NF-κB2 p100 (Ser866/870) Antibody # 4810	40 microliters	W IP	H M (R) (B) (Dg)	110	Rabbit
NF-κB2 p100/p52 Antibody # 4882	40 microliters	W IP	H M R Mk	52 active form. 120 precursor.	Rabbit
NIK Antibody # 4994	40 microliters	W IP	H M (R) (Mk) (B)	125	Rabbit
RelB Antibody # 4954	40 microliters	W IP	H M R Mk	70	Rabbit
TRAF2 Antibody # 4712	40 microliters	W	H M R Mk	53	Rabbit
TRAF3 Antibody # 4729	40 microliters	W	H M R Mk	55	Rabbit
Anti-rabbit IgG, HRP-linked Antibody # 7074	100 microliters				Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IHC-F=Immunohistochemistry (Frozen)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  B=Bovine  Dg=Dog

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

Background

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 Suppl, S81-96.
4. Xiao, G. et al. (2001) Mol Cell 7, 401-9.
5. Senftleben, U. et al. (2001) Science 293, 1495-9.
6. Xiao, G. et al. (2001) EMBO J 20, 6805-15.
7. Ling, L. et al. (1998) Proc Natl Acad Sci USA 95, 3792-7.
8. Xiao, G. et al. (2004) J Biol Chem 279, 30099-105.
9. Liang, C. et al. (2006) Cell Signal 18, 1309-17.
10. Xia, Z.P. and Chen, Z.J. (2005) Sci STKE 2005, pe7.
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.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

• 4764 NF-κB p65 (C22B4) Rabbit mAb
• 3017 NF-κB2 p100/p52 (18D10) Rabbit mAb (Human Specific)
• 3035 NF-κB1 p105/p50 Antibody
• 9936 NF-κB Pathway Sampler Kit
• 4727 c-Rel Antibody
• 3936 Ubiquitin (P4D1) Mouse mAb