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Protein Expr Purif. 2014 Apr;96:48-53. doi: 10.1016/j.pep.2014.01.014. Epub 2014 Feb 5.

A semi-automated method for purification of milligram quantities of proteins on the QIAcube.

Author information

1
Eastern New Mexico University, Department of Physical Sciences, Station #33, Portales, NM 88130, United States.
2
Eastern New Mexico University, Department of Physical Sciences, Station #33, Portales, NM 88130, United States. Electronic address: elliott.stollar@enmu.edu.

Abstract

A growing number of studies require the purification of multiple proteins simultaneously and the development of simple economical high-throughput purification methods is essential. We have tested the purification of two related proteins in a variety of conditions to benchmark the semi-automated affinity chromatography method for the QIAcube that we have developed. We find that this new QIAcube method can successfully purify milligram quantities of proteins with minimal user involvement and performs as well as methods based on gravity. The method could easily be adapted to other chromatography resins and should prove to be a versatile method for optimizing protein expression or purification conditions for multiple proteins while obtaining sufficient amounts for subsequent biochemical analyses.

KEYWORDS:

High-throughput automation; Protein expression; Protein purification; QIAcube; SH3 domain

PMID:
24508590
PMCID:
PMC4287359
DOI:
10.1016/j.pep.2014.01.014
[Indexed for MEDLINE]
Free PMC Article

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