Enzolution™ DDX41 ATPase Assay System


The Enzolution DDX41 Assay System is a comprehensive assay technology to assess the activity of DDX41, a helicase that binds and remodels viral RNA structures.

  • Enzyme, substrate, plates, and buffers
  • For use with the trusted Transcreener ADP2 Assay (Sold Separately)
  • Easy-to-use and HTS-ready
  • Ultra-sensitive detection – Use less enzyme!
SKU: 3041 Category:

A Complete Assay Solution Ready to Tackle DDX41 Discovery

The Enzolution DDX41 Assay System provides researchers with an assay tool that will simplify and advance their exploration of novel DDX41/innate immunity modulators. The assay system includes DDX41 enzyme, RNA substrate, assay plates, and buffers to generate a DDX41 enzymatic reaction. 

The Enzolution Assay System is intended for use with the trusted Transcreener ADP2 Assay (sold separately). The Transcreener ADP2 Assay is the only commercially available ADP assay that directly detects ADP production, i.e. without the use of coupling enzymes. The direct detection format minimizes compound interference, producing trustworthy results with Z' values greater than 0.7. The assay is an extensively validated ADP detection method used in millions of wells by researchers globally. 

When purchasing the Enzolution Assay System, it is important to choose the assay system readout that corresponds to the FP, FI, or TR-FRET configuration of your Transcreener ADP2 Assay.

DDX41 ATPase Assay System Schematic
DNA Damage Response and Innate Immunity Preview

Download the free eBook, The DNA Damage Response and Innate Immunity, to explore Transcreener Assays for DNA Damage Repair & Innate Immunity.

A Simple Two-Step Protocol

The DDX41 Enzyme Reaction is initiated by the addition of yeast RNA and ATP. After the Enzyme Reaction incubation is completed, ADP detection reagents are added (Transcreener ADP2 Antibody and Tracer) along with EDTA to quench the DDX41 reaction.

DDX41 Assay Outline 300 ppi

Robust HTS-Ready Assay

Z' of 0.86 confirms the robustness of the DDX41 assay. A robust system like this, is necessary to ensure the precision and efficiency required to scale-up assays for high-throughput screening.

Z' Data with DDX41 Assay

Direct Detection of DDX41 Activity with the Trusted Transcreener ADP2 Assay

Transcreener assays have fewer reagents and a less complex mechanism than any other nucleotide detection assay, which generally require coupling enzymes to convert a nucleotide to a product that can generate a signal with a reporter enzyme. The direct detection Transcreener format decreases the risk of false positives or missing a hit.

Transcreener Assay Overview

BellBrook's Recombinant DDX41 Enzyme (Included in Assay System)

Active, human full-length DDX41 protein expressed and purified from Baculovirus. For specific activity, please refer to the Certificate of Analysis for each individual enzyme lot. Purity is >70% in a composition of 50 mM Tris HCl, 200 mM NaCl, 0.5 mM TCEP, and 10% Glycerol (pH 7.5). Currently used with the Transcreener assay at a concentration of 147.4 nM in a 10 μL reaction.

DDX41 Enzyme Graphic

DDX41 Activity Under Initial Velocity Conditions

Here, we use 15 µM ATP with 0.5 mg/mL Yeast RNA substrate. The enzyme buffer included 50 mM Tris-HCl (pH 7.5), 2 mM MgCl2, and 0.01% Triton X-100. The enzyme reaction took place for 60 minutes at 30°C. The ADP Detection Mix included 1X Stop & Detect Buffer B, 4 nM ADP2 Alexa Fluor 633 Tracer, and 16 µg/mL ADP antibody. The Detection Mix was added and incubated for 60 minutes at room temperature.

Enzyme Titration - FP Readout

DDX41 Enzyme Titration with FP Readout

Linear Response

DDX41 Linear Response

Simplify Your Discovery of Novel Innate Immunity Modulators

Compounds were screened from the Tocris 2.0 Library set. An interference screen was performed to eliminate compounds interfering with detection reagents. Selected hits (SU 3327, TC LPA5 4, Polygodial, and SCH 202676) from the pilot screen were tested in dose-response mode.

Dose Response Mode

DDX41 Assay Dose Response Curve

IC50 (SU 3327) = 0.13 μM

IC50 (TC LPA5 4) = 5.36 μM

IC50 (Polygodial) = 4.43 μM

IC50 (SCH 202676) = 1.79 μM

Choose the Fluorescent Detection Mode That Best Suits Your Preference

Choose the readout configuration that is the best fit for your lab based on preference and plate reader compatibility. The assay is available with an FP, FI, or TR-FRET readout. Reference our Instrument Compatibility resource to determine the detection mode compatible with your plate reader. 

Fluorescent Intensity

DDX41 Assay Titration with FI Readout


DDX41 Assay Titration with TR-FRET Readout

Let Us Do The Work: DDX41 Assay Services

Interested in moving your DDX41 program forward, but don't want the hassle of bringing an assay in-house? BellBrook's expert scientists have you covered. Our team of biochemical and enzymology expert scientists will work diligently to progress your lead candidates through the early-stages of drug discovery. The best part - we are a small custom shop. We can customize assay conditions to suit your project needs, produce results faster, and you will directly collaborate with one of our expert scientists. 

BellBrook Scientist Performing Lead Discovery 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.
  • Residence Time Measurements Determination of koff using 'jump dilution' enzymatic assay method.
  • Mechanism of Action Studies – Kinetic analysis to define the mode of inhibition.
  • Triaging Non-stoichiometric Inhibitors – Run assays under different conditions such as varying detergent and enzyme concentrations.
  • Evaluate Compound-Target Binding – Run thermal shift assays to confirm compound-target engagement via shifts in melting temperature.

Contact a BellBrook Scientist Today

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What's Included

What You Will Need