Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Community Curation of Microbial Metabolites Enables Biological Insights of Metabolomics Data.

bioRxiv : the preprint server for biology·2026

Same author

Comparability of Liquid Chromatography Tandem Mass Spectrometry Analysis of Dissolved Organic Matter across Laboratories.

Environmental science & technology·2026

Same author

Using structured libraries, selection, and machine learning to rapidly explore the sequence space of a fluorescent deoxyribozyme.

Nucleic acids research·2025

Same author

A resource to empirically establish drug exposure records directly from untargeted metabolomics data.

Nature communications·2025

Same author

Coenzyme-protein interactions since early life.

eLife·2025

Same author

Prebiotically Plausible Peptides can Self-assemble into β-rich Nanostructures.

bioRxiv : the preprint server for biology·2025

Same journal

Aromatic Cage-Directed Azide-Methyllysine Photochemistry for Profiling Nonhistone Interacting Partners of the MeCP2 Methyl-CpG-Binding Domain.

Biochemistry·2026

Same journal

Differential Hydroxypyruvate Processing by <i>E. coli</i> and <i>P. aeruginosa</i> DXP Synthases Reveals Preferential Xylulose 5-Phosphate Formation by the <i>P. aeruginosa</i> Enzyme.

Biochemistry·2026

Same journal

Structural and Functional Characterization of Heterologous Nitrogenase Complexes.

Biochemistry·2026

Same journal

Discovery of Bacterial Unspecific Peroxygenases.

Biochemistry·2026

Same journal

Lactate Biology: Subcellular Routing and Chemical Form Define Function.

Biochemistry·2026

Same journal

Nature's Anaerobic Toolkit: Glycyl Radical Enzymes and Their Expanding Functional and Mechanistic Diversity.

Biochemistry·2026

See all related articles

Search research articles

Related Experiment Video

Updated: May 13, 2026

A Rapid and Quantitative Fluorimetric Method for Protein-Targeting Small Molecule Drug Screening

A Rapid and Quantitative Fluorimetric Method for Protein-Targeting Small Molecule Drug Screening

Published on: October 16, 2015

A Miniaturized, High-Throughput Aqueous Solvent-Centric Method for Protein Solubility Screening.

Adrian Svoboda¹, Marina Molineris¹, Theodora Tureckiova²

¹Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, Flemingovo náměstí 542/2, 160 00 Prague, Czech Republic.

|May 12, 2026

Summary

This summary is machine-generated.

This study introduces a rapid, miniaturized 96-well workflow for screening protein extraction conditions, improving soluble recombinant protein yields. The method utilizes cryogenic bead-milling and a dot-blot assay for efficient protein solubility assessment.

Keywords:

Buffer Cryomilling Heterologous expression Protein Solubility Recombinant Protein

More Related Videos

High-Throughput Screening to Obtain Crystal Hits for Protein Crystallography

High-Throughput Screening to Obtain Crystal Hits for Protein Crystallography

Published on: March 10, 2023

High-Throughput Protein Crystallization via Microdialysis

High-Throughput Protein Crystallization via Microdialysis

Published on: March 3, 2023

Related Experiment Videos

Last Updated: May 13, 2026

A Rapid and Quantitative Fluorimetric Method for Protein-Targeting Small Molecule Drug Screening

A Rapid and Quantitative Fluorimetric Method for Protein-Targeting Small Molecule Drug Screening

Published on: October 16, 2015

High-Throughput Screening to Obtain Crystal Hits for Protein Crystallography

High-Throughput Screening to Obtain Crystal Hits for Protein Crystallography

Published on: March 10, 2023

High-Throughput Protein Crystallization via Microdialysis

High-Throughput Protein Crystallization via Microdialysis

Published on: March 3, 2023

Area of Science:

Biochemistry
Molecular Biology
Protein Science

Background:

Efficiently obtaining soluble recombinant proteins is crucial for biochemical and structural studies.
Current methods often face bottlenecks in extraction and clarification processes.
Developing streamlined workflows is essential for advancing protein research.

Purpose of the Study:

To develop and validate a miniaturized 96-well workflow for screening protein extraction conditions.
To optimize the preservation of soluble recombinant protein during lysis and clarification.
To provide a rapid and cost-effective method for assessing protein solubility.

Main Methods:

Aqueous, solvent-centric, fully miniaturized 96-well workflow.
Cryogenic bead-milling of liquid-nitrogen-frozen *E. coli* pellets for uniform cell disruption.
Systematic screening of extraction conditions (pH, ionic strength, detergents, chaotropes).
Assessment of protein solubility using a 1 μL chromogenic anti-His dot-blot.

Main Results:

Demonstrated successful solubilization of challenging *de novo*-generated proteins.
Dot-blot intensity shown as a practical semiquantitative proxy for soluble protein extraction.
Provided guidelines on solvent reagent influence on downstream protein production and purification.
Workflow compatibility with upstream strategies and direct transition to scale-up.

Conclusions:

The described workflow enables efficient screening of extraction conditions for soluble recombinant protein production.
The method is rapid, cost-effective, and utilizes standard laboratory equipment.
This approach can be readily adopted or automated, accelerating protein research and development.