ALEXIS Biochemicals: RIG-I (human) (IntraCellular) Detection Set [For ELISA Application]

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ALX-746-022

Revised 11-Sep-08

Poly(deoxyadenosine:deoxythymidine) (endotoxin-free) (synthetic)
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

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

Related Products

ALX-505-008	ddWater (endotoxin-free)
ALX-505-007	PBS (endotoxin-free)
ALX-746-021	Polyinosinic-polycytidylic acid . potassium salt (TLRgrade™) (synthetic)
ALX-746-023	Oligo(deoxyadenosine:deoxythymidine) (endotoxin-free) (synthetic)

APO-54N-038

Revised 05-Dec-08

RIG-I (human) (IntraCellular) Detection Set [For ELISA Application]
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

Product Specification

SPECIES CROSSREACTIVITY:	Human
SENSITIVITY:	0.1ng/ml (range 0.156 to 10ng/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
HANDLING:	Avoid freeze/thaw cycles. After reconstitution of the standard (STD), prepare aliquots and store the reconstituted standard at -20°C.

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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

ALX-840-609	Positive Control (Cell Lysate) for Antibodies to RIG-I (human)
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ALX-210-932	Polyclonal Antibody to RIG-I (human) (AT111)