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Render Timestamp: 2024-11-21T13:22:48.317Z
Commit: 5c4accf06eb7154018ba3f54329c7590f97f534a
XML generation date: 2024-11-19 14:01:10.014
Product last modified at: 2024-11-18T12:45:19.471Z
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PDP - Template Name: Antibody Sampler Kit
PDP - Template ID: *******4a3ef3a

Rig-I Pathway Antibody Sampler Kit #8348

    Product Information

    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 two 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.

    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).
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