Immobilized metal-affinity chromatography protein-recovery screening is predictive of crystallographic structure success.

ABSTRACT: The recombinant expression of soluble proteins in Escherichia coli continues to be a major bottleneck in structural genomics. The establishment of reliable protocols for the performance of small-scale expression and solubility testing is an essential component of structural genomic pipelines. The SSGCID Protein Production Group at the University of Washington (UW-PPG) has developed a high-throughput screening (HTS) protocol for the measurement of protein recovery from immobilized metal-affinity chromatography (IMAC) which predicts successful purification of hexahistidine-tagged proteins. The protocol is based on manual transfer of samples using multichannel pipettors and 96-well plates and does not depend on the use of robotic platforms. This protocol has been applied to evaluate the expre

SUBMITTER: Choi R 

PROVIDER: S-EPMC3169392 | biostudies-literature | 2011 Sep

REPOSITORIES: biostudies-literature