BellBrook Labs
1232 Fourier Drive, Suite 115
Madison, Wisconsin 53717 USA
(608) 443-2400
info@bellbrooklabs.com
Blog
BellBrook Labs
1232 Fourier Drive, Suite 115
Madison, Wisconsin 53717 USA
(608) 443-2400
info@bellbrooklabs.com
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[Webinar] Targeting Sirtuins in Epigenetic Regulation: A High-Throughput Assay for Drug Discovery
Why HTS Campaigns Slow Down After Assay Development (and How to Keep Them Moving)
Quick Answer: Even well-developed assays can lead to delays in high-throughput screening (HTS) campaigns due to: Scaling challenges from development to screening formats Data quality issues under real screening conditions False positives that slow hit validation Workflow complexity limiting throughput Lack of clear criteria for “screen-ready” assays The most successful screening programs anticipate these challenges […]
How CROs Can Reduce Screening Costs Without Sacrificing Data Quality
Quick Answer: Contract research organizations (CROs) can reduce high-throughput screening (HTS) costs without sacrificing data quality by focusing on four key levers: Improve reagent efficiency to reduce cost per well Select assay formats that minimize interference and rework Reduce false positives to avoid costly downstream validation Simplify workflows to increase throughput and consistency The most […]
How to Optimize Kinase Assays for Z′, Sensitivity, and Cost
Quick Answer To optimize kinase assays for Z′, sensitivity, and cost, systematically tune enzyme and substrate concentrations, use ATP near Km, select an appropriate time window, and validate DMSO tolerance and controls early. Small, structured experiments—especially enzyme titration and time course—drive the biggest gains. Efficient assay formats and minimized reagent usage can reduce costs without […]
How Do You Measure Kinase Activity in High-Throughput Screening?
Quick Answer Kinase activity in high-throughput screening (HTS) is typically measured by detecting either ATP consumption, ADP production, or substrate phosphorylation. Common assay formats include ADP detection assays, phosphorylation-specific antibody assays, luminescent ATP/ADP assays, radiometric assays, mobility-shift assays, and fluorescence polarization or TR-FRET–based approaches. The best method depends on factors such as throughput requirements, susceptibility […]
Why Some Kinase Screens Produce False Positives (and How to Reduce Them)
Quick Answer: Kinase screens produce false positives when compounds interfere with assay detection, aggregate nonspecifically, undergo redox cycling, compete artificially with ATP or substrate, or disrupt assay components like luciferase. Many apparent “hits” inhibit the assay system rather than the kinase itself. Reducing false positives requires prevention-first assay design, followed by structured triage using filters, […]
Choosing Between Fluorescent and Luminescent Readouts for Kinase Assays
TL;DR Fluorescent and luminescent readouts rely on fundamentally different detection chemistries, which influence sensitivity, interference risk, workflow complexity, and cost. Luminescence is not inherently more sensitive. Well-designed direct detection fluorescent assays can be equally or even ~10× more sensitive in certain kinase contexts. Compound interference profiles differ: fluorescence is vulnerable to optical artifacts; luminescence is […]
How Genentech Used Direct Measurement of ATP Hydrolysis for the Discovery of a Clinical RIP1 Inhibitor
Drug discovery campaigns are often remembered for the final molecule that reaches the clinic. Less visible—but equally important—is the assay strategy that made the discovery possible. The identification of GDC-8264 by researchers at Genentech revealed a potent and selective RIP1 kinase inhibitor now in Phase 2 clinical development. This provides a clear example of how […]
Which HTS Assay Format Is Best for Enzyme Inhibition Studies?
Quick Answer For enzyme inhibition studies in high-throughput screening (HTS), direct detection activity assays are the preferred default primary screening format. They offer higher practical throughput, fewer interference layers, and clearer mechanistic readouts than coupled or luminescent systems. Coupled and luminescent assays remain valuable but are often best positioned as orthogonal or secondary confirmation tools. […]
Common Assay Interference Mechanisms and How to Avoid Them
Quick Answer Common assay interference mechanisms in biochemical HTS include fluorescence artifacts (auto-fluorescence and quenching), compound aggregation, redox cycling, reactive chemistry, metal chelation, detergent sensitivity, luciferase modulation, and PAINS-related liabilities. These effects can create false positives or false negatives that distort screening results. The most reliable way to avoid assay interference is to combine thoughtful […]