High-Throughput Screening Reagents: A Step-by-Step Consumables Guide for Modern Drug Discovery

High-throughput screening (HTS) enables researchers to test tens of thousands of compounds in parallel, accelerating the identification of potential therapeutics.

Whether targeting enzymes, receptors, or cellular pathways, the success of HTS depends not just on instrumentation — but on carefully selected assay reagents, microplates, and consumables engineered for precision, miniaturization, and automation.

This guide walks you through the core steps of HTS workflows, matching them with scientifically validated consumables and curated product recommendations from Avantor’s catalog. It applies equally to biochemical, phenotypic, imaging-based, and CRISPR-enabled screens.

Step 1: Biochemical HTS Assays — Building Scalable In Vitro Models

Biochemical assays are foundational to HTS. They use purified proteins or molecular targets and allow rapid, high-fidelity screening of compound libraries in solution.

Assay Formats:

Enzyme inhibition assays (e.g., kinases, proteases)
Ligand binding assays (e.g., GPCR radioligand binding)
Fluorescence or luminescence-based activity readouts
FRET/TR-FRET or AlphaScreen technologies

Critical Consumables:

Category	Scientific Role	Curated Resource
Microplates (384-/1536-well)	Miniaturize reactions while maintaining assay fidelity; choose based on detection modality (e.g., white for luminescence)	Explore Assay Plates
Detection Reagents	Fluorescent and luminescent substrates ensure sensitive signal generation from nanoliter reactions	Search Detection Kits
Reservoirs, Tips & Sealers	Precision delivery of small volumes requires automation-compatible, low-retention plastics	Automation Tips

Scientific Insight

Biochemical HTS assays often feature Z’-factors above 0.6 in well-optimized formats, making them robust for primary screens. Scaling to 1536 wells reduces reagent use by up to 80% but demands ultra-clean, low-evaporation plates and consistent compound transfer tools.

Step 2: Cell-Based HTS Assays — Bridging to Biological Relevance

Phenotypic screens conducted in living cells enable the identification of compounds that exert cellular effects beyond simple binding. These assays detect functional outcomes like cytotoxicity, pathway activation, or morphological changes.

Typical Formats:

Reporter gene assays (luciferase, GFP, β-gal)
Cell viability assays (ATP-based luminescence)
High-throughput cytotoxicity screens
GPCR or ion channel activation assays

Critical Consumables:

Category	Scientific Role	Curated Resource
Cell Culture Media & Supplements	Maintain viability and reproducibility across multi-day screens; avoid lot-to-lot variation in FBS or supplements	Browse Cell Media
TC-Treated Assay Plates	Adherent cell assays need surface-treated plates for consistent morphology and growth	TC Plates
ATP-Based Cell Viability Assays	Measure live-cell ATP as a proxy for viability (e.g., CellTiter-Glo®, ATPlite™)	Cell Viability Assays
Low Retention Pipette Tips	Reduce reagent loss during nanoliter-scale transfers	Low Retention Tips

Scientific Insight

Cell-based HTS must mitigate edge effects, evaporation, and day-to-day variability in plating. Using edge well buffers, temperature-controlled automation, and large-batch serum lots helps achieve Z’-factors >0.5.

Step 3: High-Content Screening (HCS) — Multiparametric Phenotyping with Imaging

High-content screening (HCS) combines microscopy, image analysis, and automation to quantify dozens of cellular parameters simultaneously.

Common Applications:

Subcellular localization screens (nuclear translocation, ER stress)
Multiplexed cell painting assays
Organelle-specific toxicology profiling
3D spheroid and organoid phenotyping

Critical Consumables:

Category	Scientific Role	Curated Resource
Imaging-Compatible Microplates	Black wall, optically clear bottom plates match coverslip thickness for high-resolution imaging	Search Cell Stains
Fluorescent Probes & Cell Dyes	Nuclear stains, cytoplasmic markers, live-dead dyes for multiplexed readouts	Search Cell Stains
ULA and 3D Culture Plates	Uniform spheroid formation for 3D HCS	Spheroid Plates

Scientific Insight

3D HCS assays allow modeling of in vivo-like phenotypes. Uniformity across wells (spheroid size, optical clarity) is essential for consistent image analysis. Use U-bottom or hanging drop plates for reproducibility.

Step 4: CRISPR-Based HTS — Genetic Function Screens at Scale

CRISPR-based screening enables pooled or arrayed interrogation of gene function across thousands of targets.

Use Cases:

Synthetic lethal interaction screens
Drug resistance gene identification
Reporter pathway activation via CRISPRa/CRISPRi

Critical Consumables:

Product	Description	Vendor	Link
ProDeliverIN CRISPR Kit	RNP complex delivery with high efficiency and low toxicity	OZ Biosciences	View Product
PolyMag CRISPR Kit	Magnetic-assisted plasmid/mRNA delivery of Cas9 and gRNA	OZ Biosciences	View Product
ViroMag CRISPR Kit	Lentiviral CRISPR transduction enhancer for pooled screens	OZ Biosciences	View Product
ArciTect^™ tracrRNA Kit	Synthesize tracrRNA for Cas9 editing workflows	Stemcell Technologies	View Product
NEBNext^® NGS Library Prep Kit	Pooled screen readouts by sequencing gRNA barcodes	NEB	View Product

Scientific Insight

Arrayed CRISPR HTS uses multi-well plate formats to screen hundreds of genes per plate. Pooled screens require robust cell culture formats, efficient lentiviral infection, and NGS workflows for deconvolution of enriched or depleted gRNAs.

HTS Scaling Considerations

Scaling your screen from pilot to production requires thoughtful consumables management:

Challenge	HTS Consumables Solution
Assay Miniaturization	Use 384- or 1536-well plates with validated dispensing tips
Variability	Source all reagents/plates from same lot; include intra-plate controls
Automation	Choose ANSI/SLAS-compatible plastics, barcoded plates, and stacker-ready lids
Reproducibility	Pre-qualify critical reagents, use validated protocols, and log metadata