| 货号 | 8348T |
| 描述 | The Rig-I Pathway Antibody Sampler Kit provides an economical means to evaluate the activation state and total protein levels of multiple members of the Rig-I pathway including Rig-I, MDA-5, MAVS, IRF-3, TBK1/NAK, and IKKε. The kit includes enough primary antibody to perform four western blot experiments per antibody. |
| 目标/特异性 | MDA-5 (D74E4) Rabbit mAb, Rig-I (D14G6) Rabbit mAb, MAVS Antibody, IRF-3 (D83B9) Rabbit mAb, TBK1/NAK (D1B4) Rabbit mAb, IKKε (D61F9) XP® Rabbit mAb, and IKKε (D20G4) Rabbit mAb detect endogenous levels of respective total proteins and do not cross-react with other proteins. Bands detected at 52 and 75 kDa by MAVS Antibody correlate with those described by Seth et al. (2005). Phospho-TBK1/NAK (Ser172) (D52C2) XP® Rabbit mAb detects endogenous levels of TBK1/NAK only when phosphorylated at Ser172. Phospho-IRF-3 (Ser396) (4D4G) Rabbit mAb detects endogenous levels of IRF-3 only when phosphorylated at Ser396. |
| 供应商 | CST |
| 背景 | Antiviral innate immunity depends on the combination of parallel pathways triggered by virus detecting proteins in the Toll-like receptor (TLR) family and RNA helicases, such as Rig-I (retinoic acid-inducible gene I) and MDA-5 (melanoma differentiation-associated antigen 5), which promote the transcription of type I interferons (IFN) and antiviral enzymes (1-3). TLRs and helicase proteins contain sites that recognize the molecular patterns of different virus types, including DNA, single-stranded RNA (ssRNA), double-stranded RNA (dsRNA), and glycoproteins. These antiviral proteins are found in different cell compartments; TLRs (i.e. TLR3, TLR7, TLR8, and TLR9) are expressed on endosomal membranes and helicases are localized to the cytoplasm. Rig-I expression is induced by retinoic acid, LPS, IFN, and viral infection (4,5). Both Rig-I and MDA-5 share a DExD/H-box helicase domain that detects viral dsRNA and two amino-terminal caspase recruitment domains (CARD) that are required for triggering downstream signaling (4-7). Rig-I binds both dsRNA and viral ssRNA that contains a 5-triphosphate end not seen in host RNA (8,9). Though structurally related, Rig-I and MDA-5 detect a distinct set of viruses (10,11). The CARD domain of the helicases, which is sufficient to generate signaling and IFN production, is recruited to the CARD domain of the MAVS/VISA/Cardif/IPS-1 mitochondrial protein, which triggers activation of NF-κB, TBK1/IKKε, and IRF-3/IRF-7 (12-15). |
| 存放说明 | -20C |
| 参考文献 | Yoneyama, M. and Fujita, T. (2007) J Biol Chem 282, 15315-8. Meylan, E. and Tschopp, J. (2006) Mol Cell 22, 561-9. Thompson, A.J. and Locarnini, S.A. (2007) Immunol Cell Biol 85, 435-45. Imaizumi, T. et al. (2002) Biochem Biophys Res Commun 292, 274-9. Zhang, X. et al. (2000) Microb Pathog 28, 267-78. Yoneyama, M. et al. (2005) J Immunol 175, 2851-8. Yoneyama, M. et al. (2004) Nat Immunol 5, 730-7. Hornung, V. et al. (2006) Science 314, 994-7. Pichlmair, A. et al. (2006) Science 314, 997-1001. Kato, H. et al. (2006) Nature 441, 101-5. Childs, K. et al. (2007) Virology 359, 190-200. Meylan, E. et al. (2005) Nature 437, 1167-72. Xu, L.G. et al. (2005) Mol Cell 19, 727-40. Kawai, T. et al. (2005) Nat Immunol 6, 981-8. Seth, R.B. et al. (2005) Cell 122, 669-82. |
Western blot analysis of extracts from various cell lines using IRF-3 (D6I4C) XP® Rabbit mAb. | |
Immunoprecipitation of IRF-3 from THP-1 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or IRF-3 (D6I4C) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using IRF-3 (D6I4C) XP® Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb #3678 and Anti-mouse IgG, HRP-linked Antibody #7076 were used as secondary antibodies. | |
Confocal immunofluorescent analysis of HT-29 cells, untransfected (left) or transfected with Poly(I:C) (2.5 μg/ml, 6 hr; right), using IRF-3 (D6I4C) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). | |
Western blot analysis of extracts from THP-1 cells, differentiated with TPA #4174 (80 nM, overnight) followed by treatment with LPS (1 μg/ml, indicated times), using Phospho-IKKε (Ser172) (D1B7) Rabbit mAb (upper) or IKKε (D20G4) Rabbit mAb #2905 (lower). | |
Western blot analysis of recombinant GST-IKKε and extracts from Ramos and RL7 cells using IKKε (D20G4) Rabbit mAb #2905.Western免疫印迹。用IKKε (D20G4) Rabbit mAb #2905研究Ramos 和 RL7细胞的细胞提取液。 | |
Western blot analysis of extracts from Raw 264.7 and KNRK cells using IKKε (D61F9) XP® Rabbit mAb #3416.Western免疫印迹。用IKKε (D61F9) XP® Rabbit mAb #3416研究Raw 264.7 和 KNRK 细胞的细胞提取液。 | |
Western blot analysis of extracts from THP-1 cells differentiated with TPA #4174 (80 nM, overnight) followed by treatment with LPS (1 μg/ml) for indicated times, using Phospho-TBK1/NAK (Ser172) (D52C2) XP® Rabbit mAb #5483 (upper), or TBK1/NAK (D1B4) Rabbit mAb #3504 (lower).Western免疫印迹。用Phospho-TBK1/NAK (Ser172) (D52C2) XP® Rabbit mAb #5483(上图) 或 TBK1/NAK (D1B4) Rabbit mAb #3504 (下图)研究经过 TPA #4174 (80 nM, 过夜)分化处理后又经LPS (1 μg/ml) 处理一定时间的THP-1 细胞的细胞提取液。 | |
Western blot analysis of extracts from differentiated THP-1 cells, untreated (-) or treated with LPS (1 μg/ml, 1 hr) (+), using Phospho-IRF-3 (Ser396) (4D4G) Rabbit mAb #4947 (upper) or IRF-3 (D83B9) Rabbit mAb #4302 (lower).Western免疫印迹。用Phospho-IRF-3 (Ser396) (4D4G) Rabbit mAb #4947 (上图) 或IRF-3 (D83B9) Rabbit mAb #4302 (下图)研究未经处理的 (-) 和经 LPS (1 μg/ml, 1小时) 处理的(+) 已经分化的THP-1 细胞的细胞提取液。 | |
Western blot analysis of extracts from various cell lines using IRF-3 (D83B9) Rabbit mAb #4302.Western免疫印迹。用IRF-3 (D83B9) Rabbit mAb #4302研究各种细胞系的细胞提取液。 | |
Western blot analysis of extracts from various cell lines using MAVS Antibody #3993.Western免疫印迹。用MAVS Antibody #3993研究各种细胞系的细胞提取液。 | |
Western blot analysis of extracts from differentiated THP-1 and RAW 264.7 cells, untreated (-) or LPS-treated (1 μg/ml, overnight) (+), using Rig-I (D14G6) Rabbit mAb #3743.Western免疫印迹。用Rig-I (D14G6) Rabbit mAb #3743研究未经处理的 (-) 和经 LPS (1 μg/ml, 过夜) 处理的(+) 已经分化的THP-1和RAW 264.7 细胞的细胞提取液。 | |
Western blot anlaysis of extracts from differentiated THP-1 cells, untreated or treated with LPS (1 μg/ml, indicated times), using MDA-5 (D74E4) Rabbit mAb #5321.Western免疫印迹。用MDA-5 (D74E4) Rabbit mAb #5321研究未经处理的和经 LPS (1 μg/ml)处理一定时间的已分化的THP-1细胞的细胞提取液。 | |
Flow cytometric analysis of THP-1 cells differentiated with TPA #9905, untreated (blue) or LPS-treated (green), using Phospho-TBK1/NAK (Ser172) (D52C2) XP® Rabbit mAb. | |
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with human MDA-5 (+), using MDA-5 (D74E4) Rabbit mAb. | |
Western blot anlaysis of extracts from differentiated THP-1 cells, untreated or treated with LPS (1 μg/ml, indicated times), using MDA-5 (D74E4) Rabbit mAb. |
