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

 
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2019年3月20日11時40分 現在
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NF-kB 製品一覧

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キット内容 容量 用途 種交差性 Sensitivity ホモロジー† 検出分子量 アイソタイプ
Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb #2697 20 µl WB, IHC-P, IHC-F, F H, M, R, Mk Endogenous B 85 IKK-alpha 87 IKK-beta Rabbit IgG
IKKα (3G12) Mouse mAb #11930 20 µl WB, IF-IC, F H, Mk Endogenous 85 Mouse IgG1
Phospho-NF-κB2 p100 (Ser866/870) Antibody #4810 20 µl WB, IP H, M Transfected Only R, B, Dg 110 Rabbit
NF-κB2 p100/p52 Antibody #4882 20 µl WB, IP H, M, R, Mk Endogenous 52 (mature). 120 (precursor). Rabbit
NIK Antibody #4994 20 µl WB H, M Endogenous R, Mk, B, Hr 125 Rabbit
RelB (C1E4) Rabbit mAb #4922 20 µl WB, IP H, M, R, Mk Endogenous 70 Rabbit IgG
TRAF2 Antibody #4712 20 µl WB H, M, R, Mk Endogenous 53 Rabbit
TRAF3 Antibody #4729 20 µl WB H, M, R, Mk Endogenous 62 Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl WB Endogenous Goat
Anti-mouse IgG, HRP-linked Antibody #7076 100 µl WB Horse

†Species predicted to react based on 100% sequence homology.

Applications Key: W=Western Blotting, IHC-P=Immunohistochemistry (Paraffin), IHC-F=Immunohistochemistry (Frozen), F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry), IP=Immunoprecipitation
Reactivity Key: H=Human, M=Mouse, R=Rat, Mk=Monkey

貯法
-20℃
※括弧付きの動物種は配列が100%相同であるため反応すると推定されます。
社内データ

Western Blotting

Western Blotting

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

Western Blotting

Western Blotting

Western blot analysis of extracts from THP1 and Ramos cells, using TRAF2 Antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from THP-1, L929, C6, and COS cells, using NF-kappaB2 p100 Antibody.


Western Blotting

Western Blotting

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 or total NF-κB2 p100 Antibody #4882. 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 Rutgers, The State University of New Jersey.

Western Blotting

Western Blotting

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 Blotting

Western Blotting

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


Western Blotting

Western Blotting

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

Western Blotting

Western Blotting

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

Western Blotting

Western Blotting

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


Western Blotting

Western Blotting

Western blot analysis of extacts from HeLa cells, untransfected or transfected with HA-NIK, using NIK Antibody #4994.

Western Blotting

Western Blotting

Western blot analysis of extracts from TNF-alpha and Calyculin A treated HeLa and NIH/3T3 cells, using Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb.

Western Blotting

Western Blotting

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


Flow Cytometry

Flow Cytometry

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')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human gall bladder (chronic cholecystitis), using Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb.

IF-IC

IF-IC

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).


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, untreated (left) or λ phosphatase-treated (right), using Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, showing cytoplasmic localization, using Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung (chronic bronchitis), using Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Phospho-IKKα/β (Ser176/180) (16A6) Rabbit mAb in the presence of control peptide (left) or Phospho-IKK-alpha/beta (Ser176/180) Blocking Peptide #1023 (right).

IHC-F (frozen)

IHC-F (frozen)

Immunohistochemical analysis of frozen H1650 xenograft, showing cytoplasmic localization using Phospho-IKKα/β (Ser176/180)(16A6) Rabbit mAb.

Flow Cytometry

Flow Cytometry

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.


バックグラウンド

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).

使用例
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関連製品
3017   NF-κB2 p100/p52 (18D10) Rabbit mAb
3035   NF-κB1 p105/p50 Antibody
3936   Ubiquitin (P4D1) Mouse mAb
4764   NF-κB p65 (C22B4) Rabbit mAb
4766   NF-κB Family Member Antibody Sampler Kit
4767   NF-κB p65 Antibody Sampler Kit
9936   NF-κB Pathway Sampler Kit

DRAQ5 is a registered trademark of Biostatus Limited.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
LumiGLO is a registered trademark of Kirkegaard & Perry Laboratories.
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.

本製品は試験研究用です。

NF-κB Non-Canonical Pathway Antibody Sampler Kit

Immune Cell Signaling Pathways

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