The Enzolution Assay Systems offer a complete assay solution ready to tackle your toughest enzyme targets. They include the enzyme, substrate, assay plates, and buffer components needed to perform an enzymatic reaction. The Transcreener® Assay (sold separately) detects the produced nucleotide using antibodies paired with high-affinity fluorescent tracers. Displacement of the tracer by the nucleotide causes a change in its fluorescent properties that can be read on a compatible plate reader.
BellBrook Labs focuses on developing complete assay solutions for tougher enzyme targets that are difficult to source enzymes and/or develop assays for (so researchers can focus on optimizing their lead molecules). Currently, Enzolution Assay Systems are available for:
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Overview of the Enzolution Assay System used with the Transcreener Assay.
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Example Data Using the Enzolution RIG-I Assay System
An Assay Ready to Scale for HTS
Z' data demonstrating the RIG-I assay's suitability to scale for HTS. Z' = 0.85.
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Measure Enzymatic Activity Under Initial Velocity Conditions
Enzyme reaction conditions included 100 µM ATP, 16.2 nM dsRNA, 50 mM Tris, 2.5 mM MgCl2, 50 mM NaCl, 0.01% Brij-35, and 5 mM DTT. The Detection Mix consisted of 1X Stop & Detect Buffer B, 4 nM ADP2 Alexa Fluor 633 Tracer, and 82 µg/mL ADP antibody.
RIG-I Enzyme Titration with Fluorescence Polarization (FP) Readout. The RIG-I System is available with an FP, FI, or TR-FRET readout.
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Raw polarization (mP) is converted to ADP formed using a standard curve. Conversion of raw data shows assay linearity.
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Complete Assay Solution to Screen & Profile RIG-I Inhibitors
Transcreener Assays are designed to screen compound libraries and perform follow-up dose responses in a high-throughput format.
RIG-I Dose-Response Curve with Compounds BVT 948 and Suramin. IC50 (BVT 948) = 3.56 μM, IC50 (Suramin) = 6.20 μM.
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Lead Discovery Services
BellBrook Labs also offers lead discovery services for scientists looking to advance their program without bringing a biochemical assay in-house. Their services feature custom assay conditions, quick turnaround times, and direct collaboration with a BellBrook scientist.
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Lead discovery services can include:
- Inhibitor Screening - To detect or confirm activity with the target
- Inhibitor Potency Profiling - Dose response with the target and/or associated proteins. Fast IC50 results
- Residence Time Measurements - Koff determination using the “jump dilution” method
- Mechanism of Action Studies- Kinetic study to determine the mechanism of inhibition
- Triaging Non-stoichiometric Inhibitors - Run experiments under a variety of conditions, such as varied detergent and enzyme concentrations
- Evaluate Compound-Target Binding - Using changes in melting temperature, conduct thermal shift assays to verify compound-target engagement