Aqueous Solvent-Centric Method for High-Throughput Protein Solubility Screening

Adrian Svoboda's protocol describes a miniaturized 96-well workflow for screening extraction conditions to preserve soluble recombinant protein from E. coli in a single day. The method uses cryogenic bead-milling and a 1 μL chromogenic anti-His dot-blot to assess solubility. The dataset was last updated on 2026-05-12.

Use Cases

• Optimizing protein extraction buffers based on systematic exploration of pH, ionic strength, detergents, and chaotropes.
• Screening genetic solubility-enhancement strategies using the described high-throughput workflow.
• Scaling up protein production by transitioning directly from screening hits identified by dot-blot intensity.

Strengths

• Workflow is miniaturized to a 96-well format, enabling high-throughput screening.
• Protocol is designed for completion in a single working day.
• Method uses standard molecular biology equipment and inexpensive consumables, facilitating adoption.

Limitations

• Column-level documentation is absent; field semantics must be inferred after download.
• Row count is unknown, which may limit suitability assessment.
• The dataset is small (565.9 KB), indicating limited scope, likely containing protocol documentation rather than large-scale experimental results.

Provenance

Source

Adrian Svoboda via figshare.

Collection Method

A described laboratory protocol for protein solubility screening.

Freshness

Last updated 2026-05-12 16:04:03; freshness should be verified.