Product Pathways - NF-kB Signaling
Rig-I Pathway Antibody Sampler Kit #8348
|Kit Includes||Quantity||Applications||Reactivity||MW (kDa)||Isotype|
|MDA-5 (D74E4) Rabbit mAb #5321||40 µl||W IP||H M||135||Rabbit IgG|
|Rig-I (D14G6) Rabbit mAb #3743||40 µl||W IP||H M R Mk||102||Rabbit IgG|
|MAVS Antibody #3993||40 µl||W IF-IC||H||75, 52||Rabbit|
|IRF-3 (D83B9) Rabbit mAb #4302||40 µl||W IP||H M R Mk||45-55||Rabbit IgG|
|TBK1/NAK (D1B4) Rabbit mAb #3504||40 µl||W IP||H M R Mk||84||Rabbit IgG|
|Phospho-TBK1/NAK (Ser172) (D52C2) XP® Rabbit mAb #5483||40 µl||W IP IF-IC F||H (M) (R) (Mk) (X) (B) (Dg)||84||Rabbit IgG|
|Phospho-IRF-3 (Ser396) (4D4G) Rabbit mAb #4947||40 µl||W||H M||45-55||Rabbit IgG|
|IKKε (D61F9) XP® Rabbit mAb #3416||40 µl||W IP IF-IC F||M R||80||Rabbit|
|IKKε (D20G4) Rabbit mAb #2905||40 µl||W IP||H (Mk)||80||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||Goat|
Reactivity Key: H=Human M=Mouse R=Rat Mk=Monkey X=Xenopus B=Bovine Dg=Dog
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
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.
Western blot analysis of recombinant GST-IKKε and extracts from Ramos and RL7 cells using IKKε (D20G4) Rabbit mAb #2905.
Western blot analysis of extracts from Raw 264.7 and KNRK cells using IKKε (D61F9) XP® Rabbit mAb #3416.
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 blot analysis of extracts from various cell lines using MAVS Antibody #3993.
Western blot analysis of extracts from various cell lines using IRF-3 (D83B9) Rabbit mAb #4302.
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 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.
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.
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.
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).
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For Research Use Only. Not For Use In Diagnostic Procedures.