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Antiviral Signalling / Related Products
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ALX-746-022 Revised 11-Sep-08 New product
Poly(deoxyadenosine:deoxythymidine) (endotoxin-free) (synthetic)
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SYNONYMS Poly(dA:dT) (endotoxin-free) (synthetic)
PRODUCT LINE Immunology
PRODUCT CATEGORY Antiviral Signalling / Related Products
Ordering Information
Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
ALX-746-022-C050   50 µg 320.00 USD Add To Cart
Product Specification
SOURCE/HOST: Synthetic.
QUANTITY: Aliquoted by weight of dry material. Contains polymer, residual salt and water. Sufficient for at least 200 cellular activation assays at 0.025-0.25µg/200µl.
FORMULATION: Lyophilized. Sterile.
ENDOTOXIN CONTENT: <0.02EU/µg (LAL test; BioWhittaker)
RECONSTITUTION: For a 250µg/ml stock solution, dissolve the total vial content in 200µl endotoxin-free buffer or water.
SHIPPING: AMBIENT
LONG TERM STORAGE: +4°C
USE/STABILITY: Aqueous stock solution is stable for 1 day when stored at +4°C.
HANDLING: For maximum product recovery after thawing, centrifuge the vial before opening the cap. After reconstitution, prepare aliquots and store at -20°C. Protect from light.
Product Description
Strong IFN inducer, independent of IRF3.
General Information
BACKGROUND/TECHNICAL INFORMATION

For in vitro stimulation we recommend the following procedure:
1. Bone marrow-derived mouse dendritic cells (BM mDC) are cultured 8x104/100µl in RPMI containing 10% FCS over night at 5% CO2 and 37°C.
2. The medium is removed and replaced by 100µl OPTI MEM (Invitrogen) and subsequently the transfection mixture is added: 25µl OPTI MEM containing 0.025 to 0.25 Poly(dA:dT) + 25µl OPTI MEM containing 0.5µl Lipofectamine 2000 (Invitrogen) or alternative transfection agents.
3. Over night stimulation at 5% CO2 and 37°C.
4. Measurement of IFN-α/β in culture supernatants.

General Literature References
DAI (DLM-1/ZBP1) is a cytosolic DNA sensor and an activator of innate immune response : A. Takaoka, et al.; Nature 448, 501 (2007) Abstract
Cytosolic DNA recognition for triggering innate immune responses: A. Takaoka & T. Taniguchi; Adv. Drug Deliv. Rev. 60, 847 (2008) Abstract
Intracellular pattern-recognition receptors  : C. Dostert, et al.; Adv. Drug Deliv. Rev. 60, 830 (2008) Abstract
The inflammasome recognizes cytosolic microbial and host DNA and triggers an innate immune response: D.A. Muruve, et al.; Nature 452, 103 (2008) Abstract
 
 
APO-54N-038 Revised 10-Oct-08 New product
RIG-I (human) (IntraCellular) Detection Set [For ELISA Application]
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SYNONYMS Retinoic Acid-inducible Gene 1 (human) (IntraCellular) Detection Set [For ELISA Application]
PRODUCT LINE DNA Regulation / Transcription
PRODUCT CATEGORY RNA Helicases / Related Products
Ordering Information
Product Numbers: Format: Size: Unit Price: Quantity: Add To Cart
APO-54N-038-KI01   1 Set 630.00 USD Add To Cart
Product Specification
SPECIES CROSSREACTIVITY:
Human
SENSITIVITY: 0.4ng/ml (range 0 to 32ng/ml).
QUANTITY: For 5x96 wells.
KIT/SET CONTAINS: 1 vial Standard (lyophilized) (STD).
1 vial Coating Antibody (COAT).
1 vial Detection Antibody (DET).
APPLICATION: For the quantitative determination of human RIG-I in cell extracts.
SHIPPING: SHIPPED ON BLUE ICE
SHORT TERM STORAGE: +4°C
LONG TERM STORAGE: -20°C
HANDLING: Avoid freeze/thaw cycles.
General Information
The innate immune response to viruses is initiated upon detection of viral RNA generated during the life cycle of most viruses [1]. Recognition of the viral RNA is mediated by either the two cytoplasmic RNA helicases Retinoic acid Inducible Gene-1 (RIG-I ; Ddx58) and Melanoma Differentiation Associated gene 5 (MDA5 / Helicard) [2] or by Toll-like Receptor 3 (TLR3) located to an endosomal-like compartment [3]. RIG-I and MDA5 belong to the family of RIG-I like receptor (RLR) together with the protein LGP2 whose function is still elusive [4, 5]. Upon binding to RNA, RIG-I or MDA5 binds a new mediator named Cardif, MAVS, IPS-1 or VISA [6, 7] that signals through transcription factors, such as IRF-3, IRF-7, and NF-κB leading to the activation of interferons (IFNs) and other inflammatory cytokine genes. RIG-I is activated by dsRNA and 5′-ppp RNA. RIG-I has been shown to trigger innate immune signaling pathways during infection by orthomyxoviruses, rhabdovirus, vesicular stomatitis virus, and paramyxoviruses; and possibly by the filovirus Ebola virus and by the Epstein-Barr virus [8]. Virus infection induces a rapid production of IFN-α/β that leads to expression of hundreds of interferon-stimulated genes (ISGs) whose products direct antiviral actions. RIG-I belongs to the ISG gene family and monitoring its levels using the RIG-I (human) Detection Set (IC) can be used as indicator of IFN production and virus infection.
MANUFACTURER Manufactured by Apotech Corporation.
General Literature References
[1] Recognition of viruses by innate immunity: O. Takeuchi & S. Akira; Immunol. Rev. 220, 214 (2007) Abstract
[2] MDA5/RIG-I and virus recognition: O. Takeuchi & S. Akira; Curr. Opin. Immunol. 20, 17 (2008) Abstract
[3] TLR3: Interferon induction by double-stranded RNA including poly(I:C): M. Matsumoto & T. Seya; Adv. Drug Deliv. Rev. 60, 805 (2008) Abstract
[4] Cytoplasmic recognition of RNA: M. Yoneyama, et al.; Adv. Drug Deliv. Rev. 60, 841 (2008) Abstract
[5] Loss of DExD/H box RNA helicase LGP2 manifests disparate antiviral responses: T. Venkataraman, et al.; J. Immunol. 178, 6444 (2007) Abstract; Full Text
[6] Cardif is an adaptor protein in the RIG-I antiviral pathway and is targeted by hepatitis C virus: E. Meylan, et al.; Nature 437, 1167 (2005) Abstract
[7] IPS-1, an adaptor triggering RIG-I- and Mda5-mediated type I interferon induction: T. Kawai, et al.; Nat. Immunol. 6, 981 (2005) Abstract
[8] Distinct RIG-I and MDA5 signaling by RNA viruses in innate immunity: Y.M. Loo, et al.; J. Virol. 82, 335 (2008) Abstract; Full Text
 
 

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