Rig-I Pathway Antibody Sampler Kit

• 产品编号：CST-8348S      品牌：Cell Signaling Technology       原厂货号：8348S
• 产品分类：抗体 > 一抗 > 复合抗体试剂盒
• 应用分类：

其它组分：

描述：

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 和 IKKε (D20G4) Rabbit mAb 分别能够检测对应的内源水平总蛋白，并且不与其它的蛋白交叉反应。MAVS Antibody检测到 52 和75 kDa两条条带与Seth et al. (2005)描述的有关。Phospho-TBK1/NAK (Ser172) (D52C2) XP® Rabbit mAb 只能检测内源的在Ser172位点磷酸化的TBK1/NAK蛋白。Phospho-IRF-3 (Ser396) (4D4G) Rabbit mAb只能检测内源的在Ser396位点磷酸化的IRF-3蛋白。此多克隆抗体是通过合成人源对应的MAVS C-末端周围的肽段来免疫动物而获得。抗体是通过protein A和多肽亲和层析纯化。单克隆抗体是通过合成人源对应的MDA-5 Arg470位点周围的肽段，人源对应的Rig-I Lys652位点周围的肽段，人源对应的IRF-3蛋白的C-末端肽段， 人源对应的 TBK1/NAK蛋白的Ser645位点周围肽段，鼠源对应的IKKε蛋白的C-末端肽段或者人源对应的IKKε蛋白Val345位点周围的肽段，来免疫动物而获得。活性状态的单克隆抗体是通过合成人源对应的TBK1/NAK蛋白的Ser172位点周围和IRF-3蛋白的Ser396位点周围的磷肽来免疫动物而获得。Rig-I Pathway Antibody Sampler Kit为研究Rig-I 信号通路上多个成员的活性状态和总蛋白水平提供了经济的方法，包括Rig-I, MDA-5, MAVS, IRF-3, TBK1/NAK和IKKε。本试剂盒内为每个抗体提供了足够四次Western blots 所需的一抗。天然免疫抗病毒依赖Toll样受体(TLR)家族和RNA螺旋家族中的病毒探测蛋白诱导的几条平行的信号通路的相互结合，如 Rig-I (类维生素A酸诱导的基因I) 和 MDA-5 (黑素瘤分化相关的抗原5)，它们能够促进 I 型干扰素和抗病毒酶的转录(1-3)。TLRs 和解旋酶蛋白包含识别不同病毒分子模式的位点，包括DNA、单链RNA (ssRNA)、双链RNA (dsRNA)和糖蛋白。这些抗病毒的蛋白存在于细胞的不同区域中：TLRs (i.e. TLR3, TLR7, TLR8 和 TLR9) 是在红细胞浆质膜，解旋酶定位在细胞质中。Rig-I的表达受到维生素A酸、LPS、IFN 和病毒感染的诱导 (4,5)。Rig-I和MDA-5 都包含一个检测病毒双链RNA的DExD/H盒解旋区域和两个激活下游信号通路的N-末端的caspase招募区域(CARD)(4-7)。 Rig-I能够结合到双链RNA和病毒的末端的单链RNA的5'-三磷酸上，此5'-三磷酸在寄主RNA中不存在(8,9)。虽然结构上相关，Rig-I和MDA-5能够检测不同种类的病毒(10,11)。解旋酶的CARD区域足够引起信号和IFN的产生，同时被招募到MAVS/VISA/Cardif/IPS-1线粒体蛋白的CARD区域,并引起NF-κB、TBK1/IKKε 和 IRF-3/IRF-7 的激活(12-15)。

1. Yoneyama, M. and Fujita, T. (2007) J Biol Chem 282, 15315-8.
2. Meylan, E. and Tschopp, J. (2006) Mol Cell 22, 561-9.
3. Thompson, A.J. and Locarnini, S.A. (2007) Immunol Cell Biol 85, 435-45.
4. Imaizumi, T. et al. (2002) Biochem Biophys Res Commun 292, 274-9.
5. Zhang, X. et al. (2000) Microb Pathog 28, 267-78.
6. Yoneyama, M. et al. (2005) J Immunol 175, 2851-8.
7. Yoneyama, M. et al. (2004) Nat Immunol 5, 730-7.
8. Hornung, V. et al. (2006) Science 314, 994-7.
9. Pichlmair, A. et al. (2006) Science 314, 997-1001.
10. Kato, H. et al. (2006) Nature 441, 101-5.
11. Childs, K. et al. (2007) Virology 359, 190-200.
12. Meylan, E. et al. (2005) Nature 437, 1167-72.
13. Xu, L.G. et al. (2005) Mol Cell 19, 727-40.
14. Kawai, T. et al. (2005) Nat Immunol 6, 981-8.
15. Seth, R.B. et al. (2005) Cell 122, 669-82.

原厂资料：

Specificity / Sensitivity

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.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues at the carboxy terminus of human MAVS protein. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Arg470 of human MDA-5 protein, Lys652 of human Rig-I protein, the carboxy terminus of human IRF-3 protein, Ser645 of human TBK1/NAK protein, the carboxy terminus of mouse IKKε protein, or Val345 of human IKKε protein. Activation state monoclonal antibodies are producted by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser172 of human TBK1/NAK protein or Ser396 of human IRF-3 protein.

Description

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.

Background

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).

1. Yoneyama, M. and Fujita, T. (2007) J Biol Chem 282, 15315-8.
2. Meylan, E. and Tschopp, J. (2006) Mol Cell 22, 561-9.
3. Thompson, A.J. and Locarnini, S.A. (2007) Immunol Cell Biol 85, 435-45.
4. Imaizumi, T. et al. (2002) Biochem Biophys Res Commun 292, 274-9.
5. Zhang, X. et al. (2000) Microb Pathog 28, 267-78.
6. Yoneyama, M. et al. (2005) J Immunol 175, 2851-8.
7. Yoneyama, M. et al. (2004) Nat Immunol 5, 730-7.
8. Hornung, V. et al. (2006) Science 314, 994-7.
9. Pichlmair, A. et al. (2006) Science 314, 997-1001.
10. Kato, H. et al. (2006) Nature 441, 101-5.
11. Childs, K. et al. (2007) Virology 359, 190-200.
12. Meylan, E. et al. (2005) Nature 437, 1167-72.
13. Xu, L.G. et al. (2005) Mol Cell 19, 727-40.
14. Kawai, T. et al. (2005) Nat Immunol 6, 981-8.
15. Seth, R.B. et al. (2005) Cell 122, 669-82.

注意事项：

Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150
mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02%
sodium azide. Store at –20°C. Do not aliquot the antibodies.