Heliscreener® BLM Helicase DNA Unwinding Assay Kits
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Heliscreener BLM Helicase DNA Unwinding Assay 1K

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Heliscreener Tech Manual
Single-Step DNA Unwinding Assay for Screening and Lead Discovery
  • Measure the activity of BLM helicase via DNA unwinding
  • Simple - Mix-and-read HTS-ready format
  • Uncover and validate novel BLM helicase inhibitors
  • Robust Assay Z’ > 0.7 under initial velocity conditions
  • Flexible - Use for inhibitor screening, dose-response assays, kinetic, and mechanistic analysis

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Overview – Heliscreener BLM Helicase Unwinding Assay

The Heliscreener BLM Helicase DNA Unwinding Assay is a biochemical assay designed to measure the DNA unwinding activity of BLM helicase (BLM-H). The assay utilizes a labeled double-stranded DNA substrate that is quenched when intact and emits far-red fluorescence following unwinding.

The assay can be performed in kinetic (continuous monitoring) or endpoint mode. Kinetic mode is recommended for initial velocity measurements, allowing for accurate enzyme kinetics and inhibitor IC50 determinations. Alternatively, for screening, the enzyme reaction can be stopped by adding the Stop & Detect Buffer B, and the quenched reactions can be read in endpoint mode.

This assay is designed to be used with purified enzyme preparations. Heliscreener assays are not validated with cell lysates, blood, serum, or other biological samples.

How Does Heliscreener BLM DNA Unwinding Assay Work?

Fluorescence is quenched when the Reporter DNA is intact. BLM helicase unwinds the Reporter DNA, releasing the far-red fluorescence (Cy5) from the quencher (Iowa Black® Quencher). A Capture RNA (green) hybridizes to the fluorescent strand, preventing it from re-annealing with the quenching strand.

The Heliscreener Assay is designed specifically for high-throughput screening (HTS), with a single-addition, mix-and-read format.

BLM-Helicase-Unwinding-Assay-Schematic-1030x146

It is easy to integrate into automated HTS workflows, with robust detection of DNA unwinding, and compatibility with commonly used multimode plate readers. Data quality is excellent (Z’ ≥ 0.7), and the assay uses a far-red tracer to minimize interference from fluorescent compounds and light scattering.

Far-Red FI Readouts Compatible On Major Multimode Readers

Interested in the ATPase Activity of Your Helicase?

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What’s Included

Heliscreener BLM Helicase DNA Unwinding
1K10K
ComponentPart #QtyPart #Qty
Reporter DNA, 4 μM237525 μL2376250 μL
Capture DNA, 4 μM237725 μL2378250 μL
ATP, 5 mM20531 mL20561 mL
Enzyme Assay Buffer D, 10X23132 mL231420 mL
Positive Control DNA, 4 μM237910 μL2380100 μL
DTT, 1M2296100 μL22971 mL
Stop & Detect Buffer B, 10X20271 mL203210 mL

What You Will Need

ComponentNotes
Ultrapure Nuclease Free WaterSome deionized water systems are contaminated with enzymes that can degrade both nucleotide substrates and products, reducing assay performance. Use nuclease free water such as: Invitrogen Part # AM9930
EnzymeThe BLM Helicase Heliscreener DNA Unwinding Assay is validated for use with purified BLM Helicase, which is available from BellBrook as a stand-alone product (2353, 2354).
Assay PlatesYou can purchase assay plates directly through BellBrook Labs.

It is important to use assay plates that are entirely black with a nonbinding surface. We recommend Corning® 384-well plates (Cat.# 4514). The suggested plate has a square well top that enables easier robotic pipetting and a round bottom that allows good Z’ factors. It has a recommended working volume of 15–20 µL.

Applications

  • Measure Enzymatic Activity of BLM Helicases
  • Screen Compound Libraries for BLM Helicase Inhibitors
  • Quantify Inhibitor Potency – Generate Accurate IC50s
  • Kinetic and Mechanistic Analyses

Features

  • Detection of BLM-H unwinding by DNA activity
  • Easy to use, homogenous, one-step format
  • 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 Heliscreener reagents, and read your plates. The assay is compatible with 96, 384, and 1536-well formats.

Heliscreener-BLM-Helicase-Assay-Schematic-1030x274

Tocris 2.0 Library Pilot Screen with BLM Helicase

BLM Helicase Tocris Pilot Screen

Measure Compound Inhibition with Ease

Kinetic read of BLM Helicase following the assay protocol. Measurements were taken every minute for 30 minutes using 1.5 nM BLM-H and varying amounts of the probe compound, BVT 948. The initial velocity phase of the progression curves was fitted linearly to obtain slopes that represent the enzyme’s initial rate at each compound concentration. These slopes are then plotted against compound concentration for accurate IC50 determination.

Kinetic Assay with Probe Inhibitor BVT 948

BLM Helicase Unwinding Dose Response with BVT 948

Generate Slopes From Linear Range

BLM Helicase Unwinding Dose Response with BVT 948 Linear

Plot Slopes to Obtain DRC (Kinetic)

BLM Helicase Unwinding Dose Response with BVT 948 Kinetic
IC50 = 2.676 μM

Robust HTS Ready Assay to Measure BLM-H Activity with Ease

Z’ of 0.80  or greater confirms the robustness of the Heliscreener BLM-H DNA Unwinding assay. A robust system like this is necessary to ensure the precision and efficiency required to scale up assays for high-throughput screening. No need to scale up on your own!

Heliscreener BLM Helicase Z-Prime

Assay Validated with BLM Helicase Enzyme

For screening and profiling inhibitors, choose an enzyme concentration that maximizes the signal window while still in the linear range. The assay shows outstanding sensitivity for BLM Helicase.

BLM-H

Profile BLM Helicase Inhibitors

Here are example dose-response curves with the inhibitor BVT-948. Heliscreener is readily adaptable, making dose-response assays and inhibitor profiling quick and easy.

Kinetic Mode

BLM DRC with BVT 948 in Kinetic Mode

IC50 = 2.554 μM

Endpoint Mode

BLM DRC with BVT 948 in Endpoint Mode

IC50 = 1.199 μM

DNA Helicase Assay Services

Interested in moving your DNA helicase 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.

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.
  • Inhibitor Screening – To identify or confirm activity with the target.
  • Triaging Non-stoichiometric Inhibitors – Run assays under different conditions, such as varying detergent and enzyme concentrations.
  • Evaluate Compound-Target Binding – Perform ATPase and unwinding assays to confirm target interaction.

Contact Us for Services

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The Role of DNA Helicases As Therapeutic Targets

DNA helicases are essential enzymes that regulate nearly every aspect of DNA metabolism, including replication, repair, recombination, and transcription. Their dysregulation is linked to a wide range of diseases, making them attractive therapeutic targets. In cancer, certain DNA helicases support tumor growth and genomic instability by promoting rapid DNA replication and enabling tumor cells to tolerate DNA damage.

In viral infections, both host and viral DNA helicases play key roles in viral genome replication, offering unique points of therapeutic intervention. DNA helicase dysfunction is also implicated in neurodegenerative diseases, premature aging syndromes, immunodeficiencies, and rare genetic disorders due to impaired genome maintenance and DNA damage responses.

Targeting DNA helicases with small-molecule inhibitors or nucleic acid–based therapies holds promise for developing more precise treatments across multiple disease areas.

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Contact Us to Learn More

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“Some CROs reserve their best teams for high profile customers. If you’re not right there with them you get less experienced teams… BellBrook understands enzyme kinetics, so it made that part of the project so much easier. The larger CRO didn’t seem to have the expertise we needed in this area.”

“The ADP assay is a great choice for measuring activity for any ADP-generating enzyme in both real time and endpoint formats. The universal detection system has been very useful for more challenging targets.”

“Excellent blend of simplicity & good science. Excellent technical support”

“The assay itself has good reproducibility and provides a clear readout on our enzyme activity. The most important piece, however, is the level of attention to our project and rapid turnaround time. The staff at BellBrook knows what they are doing.”

“The Transcreener AMP/GMP Assay is extremely straightforward to use and is very sensitive. In a UV-based enzyme assay I needed 1 µM enzyme to see a decent signal; however using the Transcreener AMP assay I only need 10 nM. Also, BellBrook Labs have been very helpful with the assay development. Their representative has been very willing to quickly help me get my assay up and running.”

“Through the process of developing and validating an assay for HTS applications, I generated a very nice data package of robust high quality data.”

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BellBrook Labs
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Madison, Wisconsin 53717 USA
(608) 443-2400

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