Targeting OAS Enzymes with the Transcreener 2-5A OAS Assay

OAS Assay Method

Ordering Information

Assay Services

Related Resources

Transcreener 2-5A OAS Assay

Transcreener OAS Assay measures the 2-5A produced by OAS1 (2'-5'-oligoadenylate synthetase 1) OAS2, or OAS3. These measurements allow researchers to effectively determine the activity of the OAS enzyme. The assay provides a powerful tool to screen compound libraries for OAS modulators to help find new therapies for disease.

The kit comes complete with the detection reagents required to measure activity. OAS enzyme is not included in the inhibitor screening assay kit. Please contact us for questions related to acquiring the enzyme.

How Does This OAS Assay Work?

The Transcreener OAS Assay determines OAS enzyme activity by using a coupling enzyme that converts 2'-5'A produced by OAS1 to the products AMP and ATP. AMP can be measured using the trusted Transcreener AMP² Assay components. The assay provides a universal method to assess activity of any AMP-producing enzyme.

Transcreener OAS Assay technology uses a simple but highly effective method that consists of an antibody selective to AMP and a far-red fluorescent tracer. AMP produced in the reaction competes with the tracer changing the fluorescent properties and providing fluorescent readout.

The OAS assay is available with FP readout. It is a simple mix-and-read format. Perform your enzyme reaction, add the detection reagent, and measure. The simplicity of the system yields robust results that also makes it extremely amiable to HTS.

Detection of 2-5A to Measure OAS Enzymatic Activity

Fluorescent Polarization (FP)

In the presence of dsRNA OAS uses ATP to produce the messenger 2-5A. The assay uses a coupling enzyme to generate AMP from 2-5A. AMP is measured by the Transcreener AMP/GMP Assay.

Learn More About The FP Assay

Transcreener OAS FP Assay Tech Manual

Applications

Measure Enzymatic Activity of OAS1, OAS2, or OAS3
Screen Compound Libraries for OAS Modulators
Quantify Inhibitor Potency
Inhibitor Selectivity Profiling

Features

Designed as in homogenous, one-step format to enable reproducibility.
Robust Assay Z’ > 0.7 under initial velocity conditions
Far-red fluorescent readouts minimize compound interference

Easy-to-Use, Mix-and-Read, HTS-Ready Assay

Run your enzyme reaction, add Transcreener reagents, and read your plates. The OAS assay is compatible with 96, 384, and 1536-well formats.

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Robust Assay Yields Quality Data

Z’ measurements using optimized OAS1 reaction conditions indicate a robust assay. Robust data like this is vital for sizeable high throughput screens that are difficult to complete due to the massive sample quantity. Z' shown here = 0.87.

Enzyme Titration with the Transcreener OAS Assay - FP Readout

The assay demonstrates linearity when raw data is converted to the product formed using a standard curve. Linearity is shown under initial velocity conditions when raw data is converted to AMP formed as 2-5A is broken down by the detection reagents.

Human OAS1 Enzyme Titration

Linear Response

Screen for Inhibitors & Perform SAR

Transcreener Assays are designed for screening compound libraries in a high throughput format. Follow-up SAR can also be performed using the assay to determine inhibitor potency with ease.

Dose-Response with Suramin

IC₅₀ = 0.162 µM

BellBrook Lab's Recombinant OAS1 Enzyme

Active, full-length human 2'-5' oligoadenylate synthetase 1 enzyme, His-tagged protein expressed and purified from E.coli. The enzyme has been thoroughly validated with Transcreener 2-5A OAS Activity Assays. For specific activity, please refer to the Certificate of Analysis for each individual enzyme lot. Purity is >95% in a 20 mM Tris-HCl pH 8.0, 10% glycerol, 1 mM DTT buffer composition. Currently used with the Transcreener assay at a concentration between 3 and 16 nM.

Go to Human OAS1 Enzyme Page

OAS1 Assay Services

Interested in moving your program forward, but don't want to bring an assay in-house? Our scientists can help! BellBrook scientists will use their extensive biochemistry and enzymology expertise to work with you and accelerate your OAS1 or innate immunity program.

Lead Discovery Services Include:

Inhibitor Screening – To identify or confirm activity with the target.
Inhibitor Potency Profiling – Dose-response with target and/or related proteins. Fast IC₅₀ results.
Mechanism of Action Studies – Kinetic analysis to define the mode of inhibition.

Contact Us to Learn More

Please fill out the form below. We will respond quickly to get the conversation moving and learn how we can help. We keep things discrete, confidential, and professional.

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Far-Red FP Readout Validated on Major Multimode Readers

Supplier	Instrument	FP Assays
	TriStar²S LB 942	in review
	Mithras² LB 943	in review

	Cytation™ 5	validated
	Cytation™ 3	validated
	Cytation™ 1	validated
	Synergy™ H1	validated
	Synergy™ 2/H4/4	validated
	Synergy™ HTX	not capable
	Synergy™ Neo 2	validated

	POLARstar^® Omega	validated
	FLUOstar^® Omega	not capable
	PHERAstar^® FSX	validated
	PHERAstar^® Plus/FS	validated
	CLARIOstar^® /Plus	validated
	VANTAstar^™	validated

	Analyst^® GT/HT	validated
	Gemini^® XPS/EM	not capable
	SpectraMax^® M2/M2e	not capable
	SpectraMax^® M5/M5e/FlexStation^® 3	not capable
	SpectraMax^® Paradigm	validated

	EnVision^®/EnVision^® Xcite	validated

	Infinite^® M1000/M1000Pro/Safire2™	validated
	Infinite^® M200	not capable
	Infinite^® F500	validated
	Infinite^® F200/Ultra^evolution	contact us
	Spark™ 10M	validated

What's Included

Component	Notes
AMP² Antibody	Antibody specific to the AMP produced by the 2-5A Detection Mix.
AMP Tracer	When displaced, changes in fluorescence can be read in an FP assay format.
dsRNA	Used to activate OAS enzymes.
AMP	AMP is used to create the ATP/AMP standard curve.
ATP	ATP is used to create the ATP/AMP standard curve and as a substrate for OAS.
2-5A Coupling Enzyme	Transfers 2-5A to AMP allowing measurement of AMP by the Transcreener AMP² Assay components.
OAS Buffer, 10X	Buffer prepared for use with OAS enzymes. 250 mM Tris, 100 mM MgCl₂, 0.1% Brij-35.

What You Will Need

Component	Notes
OAS Enzyme	We have successfully used purified recombinant OAS1 available through BellBrook Labs. Cat. #2249 at a concentration between 3 and 16 nM. Please titrate your enzyme as stated in the technical manual for best results.
Assay Plates	Assay Plates Can Be Purchased Directly Through BellBrook Labs. FP - An entirely black plate with a non-binding surface is required. We suggest Corning 384-well plates Cat. #4514
Plate Reader	A multi-detection microplate reader configured to measure the output of the AMP tracer is required.
Liquid Handling Devices	Use liquid handling devices that can accurately dispense sub-microliter volumes into 384-well plates.
Ultrapure Water	Some deionized water systems are contaminated with nucleases that can degrade both nucleotide substrates and products, reducing assay performance. Careful handling and use of ultrapure water eliminates this potential problem.

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Transcreener OAS Assay Kits

* For custom or bulk orders (over 10,000 wells) please contact us (info@bellbrooklabs.com) for a quote.

Cat. No.	Product Description	Price
3027-1K	Transcreener OAS FP Assay (1,000 Assay, 384 Well)	$995	Add to Cart
3027-10K	Transcreener OAS FP Assay (10,000 Assay, 384 Well)	$5,699	Add to Cart

Human OAS1 Enzyme

* For custom or bulk orders (over 10 μg) please contact us (info@bellbrooklabs.com) for a quote.

Cat. No.	Product Description	Price	Add to Cart
2249	Recombinant OAS1 Enzyme, Active, Human, Full-Length, 10 µg	$285.00	Add to Cart

Assay Plates for FP

Cat. No.	Product Description	Price
2233	Corning 96-Well Black Assay Plates (#3686), 3-Pack (200+ Assays)	$50.00	Add to Cart
2229	Corning 384-Well Black Assay Plates (#4514), 3-Pack (1,000+ Assays)	$55.00	Add to Cart
2230	Corning 384-Well Black Assay Plates (#4514), 30-Pack (10,000+ Assays)	$449.00	Add to Cart

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The Role of OAS1 As a Therapeutic Target

Double-stranded RNA enters the cell from viruses. As a first line of defense, viral double-stranded RNA activates OAS1. OAS1 then synthesizes 2-5A from ATP, which activates RNase L. This signaling cleaves both viral and host RNA leading to inhibition of protein synthesis. The replication of invading RNA viruses ends.

Mutation or inadvertent stimulation of OAS1 can produce autoimmune conditions by activating the pathway inconsistently, even while not in the presence of viral invaders. Because of this, OAS1 is an interesting target in drug discovery.

The Transcreener OAS Assay is an excellent tool for researchers examining the therapeutic effects of OAS1.

Transcreener® OAS Assay

Targeting OAS Enzymes with the Transcreener 2-5A OAS Assay

OAS Assay Method

Ordering Information

Assay Services

Related Resources

Transcreener 2-5A OAS Assay

How Does This OAS Assay Work?

Detection of 2-5A to Measure OAS Enzymatic Activity

Fluorescent Polarization (FP)

Applications

Features

Easy-to-Use, Mix-and-Read, HTS-Ready Assay

Robust Assay Yields Quality Data

Enzyme Titration with the Transcreener OAS Assay - FP Readout

Human OAS1 Enzyme Titration

Linear Response

Screen for Inhibitors & Perform SAR

Dose-Response with Suramin

IC50 = 0.162 µM

BellBrook Lab's Recombinant OAS1 Enzyme

OAS1 Assay Services

Lead Discovery Services Include:

Contact Us to Learn More

Far-Red FP Readout Validated on Major Multimode Readers

What's Included

What You Will Need

Transcreener OAS Assay Kits

Human OAS1 Enzyme

Assay Plates for FP

The Role of OAS1 As a Therapeutic Target

Related Resources

IC₅₀ = 0.162 µM