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

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)

Transcreener OAS1 Assay Schematic

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.

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. 

Transcreener Mix and Read Assay

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

OAS1 Assay Enzyme Titration Raw Data

Linear Response

Linear Response Data OAS Assay

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

OAS1 Assay Dose Response 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.

OAS1 Enzyme

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.

Assay Development Services

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.

Far-Red FP Readout Validated on Major Multimode Readers

Supplier Instrument FP Assays
bioteklogo TriStar²S LB 942 in review
Mithras² LB 943 in review
bioteklogo 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
BMGLABTECH Logo POLARstar® Omega validated
FLUOstar® Omega not capable
PHERAstar® FSX validated
PHERAstar® Plus/FS validated
CLARIOstar® /Plus validated
VANTAstar validated
MDS AT logo Analyst® GT/HT validated
Gemini® XPS/EM not capable
SpectraMax® M2/M2e not capable
SpectraMax® M5/M5e/FlexStation® 3 not capable
SpectraMax® Paradigm validated
perkinElmerLogo EnVision®/EnVision® Xcite validated
tecanLogo Infinite® M1000/M1000Pro/Safire2™ validated
Infinite® M200 not capable
Infinite® F500 validated
Infinite® F200/Ultraevolution contact us
Spark™ 10M validated

What's Included

What You Will Need

Transcreener OAS Assay Kits

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

Human OAS1 Enzyme

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

Assay Plates for FP

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.

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