| 货号 | 9936T |
| 描述 | 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. |
| 目标/特异性 | 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. |
| 供应商 | CST |
| 背景 | 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. |
| 存放说明 | -20C |
| 参考文献 | Yamamoto, Y. and Gaynor, R.B. (2004) Trends Biochem. Sci. 29, 72-79. Ghosh, S. and Karin, M. (2002) Cell 109, S81-S96. Viatour, P. et al. (2005) Trends Biochem. Sci. 30, 43-52. |
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. | |
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 various cell lines using IKKα (3G12) Mouse mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). | |
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 HeLa, NIH/3T3 and PC12 cells, using IκB-α (L35A5) Mouse mAb (Amino-terminal Antigen) #4814. | |
Western blot analysis of extracts from HeLa and NIH/3T3 cells, untreated or treated with TNF-α (#2169, 20 ng/ml) for 5 minutes, using Phospho-IκB-α (Ser32) (14D4) Rabbit mAb #2859 (upper), or IκBα (44D4) Rabbit mAb #4812 (lower). | |
Western blot analysis of extracts from HeLa and NIH/3T3 cells, untreated or TNF-α treated (#2169, 20 ng/ml for 5 minutes), using Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb #3033 (upper) or NF-κB p65 Antibody #3034 (lower). | |
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 #2697. | |
Western blot analysis of extracts from HeLa, NIH/3T3 and PC-12 cells using IIKKβ (2C8) Rabbit mAb #2370. | |
Western blot analysis of extracts from NIH/3T3, HeLa and PC12 cells, using IKKα Antibody #2682. | |
Western blot analysis of extracts from various cell lines using IKKβ (D30C6) Rabbit mAb. | |
Western blot analysis of extracts from wild-type, IKKα (-/-), and IKKβ (-/-) mouse embryonic fibroblasts (MEFs) using IKKβ (D30C6) Rabbit mAb (upper) and GAPDH (14C10) Rabbit mAb #2118 (lower). | |
Western blot analysis of extracts from various cell lines using NF-κB p65 (D14E12) XP® Rabbit mAb #8242. | |
Immunohistochemical analysis of paraffin-embedded human chronic cholecystitis using NF-κB p65 (D14E12) XP® Rabbit mAb. | |
Flow cytometric analysis of HeLa cells using NF-κB p65 (D14E12) XP® Rabbit mAb (blue) compared to concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). |
