NF-κB Non-Canonical Pathway Antibody Sampler Kit【科瑞奥博】

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NF-κB Non-Canonical Pathway Antibody Sampler Kit

货号： 4888T 基本售价： 6830.0 元规格： -

产品信息

概述
货号	4888T
描述	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.
目标/特异性	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.

性能
供应商	CST
背景	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).
存放说明	-20C
参考文献	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 U S A 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.

参考图片

Flow cytometric analysis of THP-1 cells, untreated (blue) and with TPA and LPS (green) using IKK-α (Ser176/Ser180) phosphate Rabbit mAb. Anti-rabbit IgG (H+L), F(ab)2 Fragment (PE Conjugate) #8885 was used as a secondary antibody.

Western blot analysis of extracts from various cell lines using IKKα (3G12) Mouse mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Flow cytometric analysis of HCT 116 cells using IKKα (3G12) Mouse mAb (blue) compared to concentration-matched Mouse (G3A1) mAb IgG1 Isotype Control #5415 (red). Anti-mouse IgG (H+L), F(ab)₂Fragment (Alexa Fluor^® 488 Conjugate) #4408 was used as a secondary antibody.

Western blot analysis of extracts from Raji, THP-1 and BaF3 cells using RelB (C1E4) Rabbit mAb.

Western blot analysis of extracts from COS-7 cells, untransfected (-) or transfected with human TRAF3 construct (+) using TRAF3 Antibody.

Western blot analysis of extracts from various cell lines using TRAF3 Antibody.

Western blot analysis of extracts from various cell lines, untreated or treated with 10uM MG132, using NIK Antibody #4994.

Confocal immunofluorescent analysis of HCT 116 (high expression; left) and IGROV-1 (low expression; right) cells using IKKα (3G12) Mouse mAb (green). Blue pseudocolor = DRAQ5^® #4084 (fluorescent DNA dye).

Western blot analysis of extracts from THP-1 cells, differentiated with TPA (#9905, 80 nM for 24h) and treated with 1 μg/ml LPS for the indicated times, using Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb.

Western blot analysis of extracts from HeLa cells transfected with wild-type or mutant NF-κB2 p100 (SS866/870AA) and with or without NIK, using Phospho-NF-κB2 p100 (Ser866/870) Antibody #4810 (upper) and total NF-κB2 p100/p52 Antibody #4882 (lower). The p100 constructs were generously provided by Dr. Warner Greene of the Gladstone Institute of Virology and Immunology, Dr. Shao-Cong Sun of The Pennsylvania State University College of Medicine, and Dr. Gutian Xiao of the University of Pittsburgh Medical Center.Western免疫印迹。用Phospho-NF-κB2 p100 (Ser866/870) Antibody #4810 (上图)和total NF-κB2 p100/p52 Antibody #4882 (下图)研究转染了野生型和突变型NF-κB2 p100 (SS866/870AA)，具有NIK或者没有NIK的HeLa细胞的细胞提取液。p100质粒由Dr. Warner Greene of the Gladstone Institute of Virology and Immunology, Dr. Shao-Cong Sun of The Pennsylvania State University College of Medicine, and Dr. Gutian Xiao of the University of Pittsburgh Medical Center友好提供。

Western blot analysis of extracts from NIH/3T3, HeLa and PC12 cells using IKKα Antibody #2682.Western免疫印迹。用IKKα Antibody #2682研究NIH/3T3, HeLa 和 PC12 细胞的细胞提取液。

Western blot analysis of extracts from TNF-α (#2169, 20 ng/ml) and Calyculin A (#9902, 50 nM) treated HeLa and NIH/3T3 cells, using Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb #2697.Western免疫印迹。用Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb #2697研究经 TNF-α (#2169, 20 ng/ml) 和 Calyculin A (#9902, 50 nM) 处理的HeLa和NIH/3T3 细胞的细胞提取液。

Western blot analysis of extracts from Raji (human), PC12 (rat) and C2C12 (mouse) cell lines using TRAF3 Antibody #4729.Western免疫印迹。用TRAF3 Antibody #4729研究Raji (人源), PC12 (大鼠源) 和 C2C12（小鼠源）细胞系的细胞提取液。

Western blot analysis of extacts from HeLa cells, untransfected or transfected with HA-NIK, using NIK Antibody #4994.Western免疫印迹。用NIK Antibody #4994研究未转染的和转染了HA-NIK的HeLa细胞的细胞提取液。