Sensitive EdU Cell Proliferation Assay for High-Throughput Screening

Product Information

High-throughput DNA synthesis detection with minimal cytotoxicity and automation-ready workflow

ClickTech Sensitive EdU Cell Proliferation Kit for HTS is engineered for large-scale screening in 96-well and higher-density formats, delivering precise, reproducible quantification of cell proliferation for drug discovery, toxicity profiling, and compound libraries. The assay uses EdU (5-ethynyl-2′-deoxyuridine) incorporation during DNA synthesis and a bioorthogonal click reaction with picolyl-azide fluorophores, ensuring bright, stable signals and minimal background, ideal for high-content analysis across diverse cell types.

Automation Compatibility

• Fully compatible with liquid handling robots and automated plate washers
• Optimized for automated reagent dispensing and high-content screening platforms
• Seamless integration with LIMS and data analysis pipelines
• Reagent volumes and incubation times tailored for robotic scheduling
• Minimal manual intervention, ideal for unattended batch processing

Key Benefits

• Exceptional Sensitivity with low background: Optimized labeling chemistry and picolyl‑azide fluorophore deliver high signal‑to‑noise even in sparse wells (≈ ≥ 500 cells/well) and low‑proliferation conditions.
• Non-Radioactive & Safer: Eliminates hazards of [³H]-thymidine and BrdU; no harsh treatments or specialized equipment.
• Fast, Simplified Workflow: Complete the assay in under 2 hours, no DNA denaturation or antibody steps; fully automation-ready.
• Low Copper, High Viability: The click step is formulated with reduced copper versus conventional EdU protocols, supporting cell health during longer incubations and preserving compatibility with downstream readouts.
• Automation-Ready: Assay timings, wash steps, and reagent additions are tuned for liquid handlers, automated plate washers, and unattended batch processing, with consistent performance across plates, days, and operators.
• Scalable & Reproducible: Bioorthogonal chemistry supports thousands of wells with consistent performance across plates and runs.

Recommended workflow (plate‑ready)

1. Cell preparation & EdU pulse: Seed cells in black, clear‑bottom plates. Pulse with EdU during the intended window (e.g., 15–120 min). Short pulses capture S‑phase entry; longer pulses give cumulative labeling for slow cyclers.
2. Fix & permeabilize: Apply standard aldehyde fixation followed by mild permeabilization to retain morphology and antigenicity for imaging or multiplex staining.
3. Click detection: Add the picolyl‑azide 488 dye cocktail. Incubate under gentle conditions, wash.
4. Optional multiplex: Introduce viability dyes, phenotypic markers, or nuclear stains (for HCS segmentation) as needed.
5. Readout:

• Plate reader: bottom read with 488‑compatible excitation and 516 nm emission filter.
• HCS/Imaging: use the green channel; quantify EdU‑positive nuclei or integrated intensity per cell.

Plate normalization tip: include row/column controls to support ratio‑based normalization and drift correction; monitor Z′‑factor and well‑to‑well CV before production runs.

Applications

• High-throughput proliferation and cell-cycle screening
• Genotoxicity and mechanism-of-action studies
• Anticancer compound profiling and SAR workflows
• Stem cell proliferation and differentiation monitoring
• Orthogonal readouts with viability/apoptosis markers or HCS imaging

LITERATURE

EdU Incorporation To Assess Cell Proliferation and Drug Susceptibility in Naegleria fowleri. 2021, Antimicrob Agents Chemother.

https://doi.org/1128/AAC.00017-21

Multiplexed and reproducible high content screening of live and fixed cells using Dye Drop. Nature Communications. 2022.

https://doi.org/10.1038/s41467-022-34536-7

Efficient Tandem Copper‐Catalyzed Click Synthesis of Multisugar‐Modified Oligonucleotides. Angewandte Chemie International Edition, e202405161.

https://doi.org/10.1002/anie.202405161