2, 5-Oligoadenylate Synthetase ELISA Kit (2, 5-OAS)

Full Name: 2, 5-Oligoadenylate Synthetase ELISA Kit (2, 5-OAS)
Reactivity: Human
Sample Type: Plasma, Tissue Homogenates, Serum, Biological Fluids
Sensitivity: 0.281 ng/ml

INTRODUCTION

2′,5′-oligoadenylate synthetase (OAS) refers to a family of interferon-inducible enzymes that play an integral role in the innate immune response against viral infections. Upon detection of double-stranded RNA intermediates produced during viral replication, OAS proteins activate RNase L to degrade viral and cellular RNAs, potently inhibiting viral propagation. The OAS family includes multiple isozymes encoded by separate genes, which contain conserved synthetase domains but differ in their activation requirements and kinetics.

Among the OAS family, OAS1 encodes the p42 and p46 isoforms, which represent the major synthetases producing 2’,5’-oligoadenylates (2-5A) – unusual 2’-5’ phosphodiester-linked oligomers of adenosine. Binding of double-stranded RNA to the OAS1 catalytic groove results in a conformational change, leading to activation of its synthetase activity. Subsequently, OAS1 converts ATP into pyrophosphate and 2-5A molecules ranging from dimers to tetramers. These 2-5As then bind to and activate latent RNase L, which degrades single-stranded viral and cellular RNAs, thereby crippling viral infection. Additionally, OAS proteins exert antiviral activity through RNAse L-independent mechanisms.

Aberrant OAS activity has been linked to several inflammatory disorders and susceptibility to certain viruses such as West Nile and Dengue. Moreover, upregulation of RNAse L activation by higher interferon levels may promote autoimmune conditions like systemic lupus erythematosus. Hence, tight regulation of OAS-RNAse L signaling is imperative, and mechanisms that selectively enhance antiviral activity over detrimental inflammatory effects are of high interest. Recent structural studies highlight promising opportunities to modulate OAS1 activity through allosteric regulation, post-translational modifications, alternative splicing, and targeting of protein-RNA interactions governing activation.

INTENDED USE

Human 2, 5-oligoadenylate synthetase ELISA kit can measure concentrations of 2, 5-OAS (2, 5-oligoadenylate synthetase, 2-5A synthase 1, E18/E16, p46/p42 OAS, OIAS, OAS1) present is plasma, serum, biological fluids or tissue homogenate samples.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

One 96-Well Plate: Pre-coated with anti-2, 5-OAS antibody.
Standards: Lyophilized recombinant.
Sample/Standard Dilution Buffer.
Biotinylated-labelled Antibody.
Antibody Dilution Buffer.
HRP-Streptavidin Conjugate (SABC).
SABC Dilution Buffer.
TMB Substrate.
Wash Buffer (25x).
Plate Sealer.
Product Instructions.

TYPICAL RESULTS

For this 2, 5-oligoadenylate synthetase ELISA kit, it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 0.469, 0.938, 1.875, 3.75, 7.5, 15, 30 ng/ml.
Reactivity: Human
Sensitivity: 0.281 ng/ml
Range: 0.469 – 30 ng/ml
Principle: Sandwich, Double Antibody
Application: Research Use Only.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for 2, 5-OAS, no cross reactivity or interference between 2, 5-OAS and analogues was detected.
– Recovery: Serum (85 – 101%), EDTA Plasma (89 – 101%), Heparin Plasma (86 – 104%).
– Linearity: Serum (85 – 100%), EDTA Plasma (91 – 100%), Heparin Plasma (80 – 90%).
– Precison Intra-Assay: CV < 8%
– Precison Inter-Assay: CV < 10%

REFERENCES

Gene structure and function of the 2′-5′-oligoadenylate synthetase family. Cell Mol Life Sci. (2000) 57 (11): 1593-612. Justesen J., et al.
SOCS-1 and SOCS-3 inhibit IFN-alpha-induced expression of the antiviral proteins 2,5-OAS and MxA. Biochem Biophys Res Commun. (2004) 320 (3): 1007-14. Vlotides G., et al.
In vivo treatment of HCV core-positive HepG2 cells with the transfer of recombinant caspase-3 using a 2′-5′ OAS promoter. Mol Med Rep. (2012) 5 (3): 631-6. Zi Y., et al.
Effects of specific mutations in active site motifs of 2′,5′-oligoadenylate synthetase on enzymatic activity. J Interferon Cytokine Res. (2000) 20 (3): 337-44. Yamamoto Y., et al.

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