Explanatory chapter: troubleshooting protein expression: what to do when the protein is not soluble

Krisna C Duong-Ly; Sandra B Gabelli

doi:10.1016/B978-0-12-420119-4.00018-5

Explanatory chapter: troubleshooting protein expression: what to do when the protein is not soluble

Methods Enzymol. 2014:541:231-47. doi: 10.1016/B978-0-12-420119-4.00018-5.

Authors

Krisna C Duong-Ly¹, Sandra B Gabelli²

Affiliations

¹ Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
² Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Electronic address: gabelli@jhmi.edu.

PMID: 24674075
DOI: 10.1016/B978-0-12-420119-4.00018-5

Abstract

Production of soluble protein remains a bottleneck in the biochemistry and structural biology fields. Unfortunately, there is no 'magic bullet' that solves all solubility problems. The following is a protocol to test whether a protein expressed recombinantly is soluble, and possible strategies to circumvent insolubility issues.

Keywords: Cell lysis; Differential scanning fluorimetry (DSF); IPTG; Immunoblotting; SDS-PAGE; Small ubiquitin-like modifier (SUMO); Troubleshooting protein expression.

MeSH terms

Protein Engineering / methods*
Recombinant Proteins / chemistry*
Solubility

Substances

Recombinant Proteins