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8348S 1 Kit (9 x 40 µl) $559.00
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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
MDA-5 (D74E4) Rabbit mAb 5321 40 µl
H M 135 Rabbit IgG
Rig-I (D14G6) Rabbit mAb 3743 40 µl
H M R Mk 102 Rabbit IgG
MAVS Antibody 3993 40 µl
H 75, 52 Rabbit 
IRF-3 (D6I4C) XP® Rabbit mAb 11904 40 µl
H Mk 50-55 Rabbit IgG
TBK1/NAK (D1B4) Rabbit mAb 3504 40 µl
H M R Mk 84 Rabbit IgG
Phospho-TBK1/NAK (Ser172) (D52C2) XP® Rabbit mAb 5483 40 µl
H 84 Rabbit IgG
Phospho-IRF-3 (Ser396) (4D4G) Rabbit mAb 4947 40 µl
H M 45-55 Rabbit IgG
Phospho-IKKε (Ser172) (D1B7) Rabbit mAb 8766 40 µl
H 80 Rabbit IgG
IKKε (D20G4) Rabbit mAb 2905 40 µl
H 80 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
All Goat 

Product 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.

Specificity / Sensitivity

MDA-5 (D74E4) Rabbit mAb, Rig-I (D14G6) Rabbit mAb, MAVS Antibody, IRF-3 (D6I4C) XP® Rabbit mAb, TBK1/NAK (D1B4) 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. This antibody may cross-react with phospho-IKKε. Phospho-IRF-3 (Ser396) (4D4G) Rabbit mAb detects endogenous levels of IRF-3 only when phosphorylated at Ser396. Phospho-IKKε (Ser172) (D1B7) Rabbit mAb recognizes endogenous levels of IKKε protein only when phosphorylated at Ser172. This antibody may cross-react with phospho-TBK1/NAK.

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, Ser645 of human TBK1/NAK protein, Val345 of human IKKε protein, or recombinant human IRF-3 protein. Activation state monoclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser172 of human TBK1/NAK protein, Ser396 of human IRF-3 protein, or Ser172 of human IKKε protein.

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.

Entrez-Gene Id 9641
Swiss-Prot Acc. Q14164

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 5,675,063.