Skip to content

Preparation and extraction of insoluble (Inclusion-body) proteins from Escherichia coli

by Ira Palmer, Paul T. Wingfield
Current Protocols in Protein Science ()
Get full text at journal


High-level expression of many recombinant proteins in Escherichia coli leads to the formation of highly aggregated protein commonly referred to as inclusion bodies. Inclusion bodies are normally formed in the cytoplasm; alternatively, if a secretion vector is used, they can form in the periplasmic space. Inclusion bodies can be recovered from cell lysates and this unit describes preparation of washed pellets and solubilization of the protein using guanidine x HCl. The extracted protein, which is unfolded, is either directly folded as described in UNIT or further purified by gel filtration in the presence of guanidine x HCl as idescribed here. A support protocol describes the removal of guanidine x HCl from column fractions so they can be monitored by SDS-PAGE.

Cite this document (BETA)

Readership Statistics

454 Readers on Mendeley
by Discipline
71% Agricultural and Biological Sciences
11% Biochemistry, Genetics and Molecular Biology
9% Chemistry
by Academic Status
35% Student > Ph. D. Student
17% Researcher
15% Student > Master
by Country
1% United Kingdom
1% United States
1% India

Sign up today - FREE

Mendeley saves you time finding and organizing research. Learn more

  • All your research in one place
  • Add and import papers easily
  • Access it anywhere, anytime

Start using Mendeley in seconds!

Sign up & Download

Already have an account? Sign in