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GST-His purification: A Two-step Affinity Purification Protocol Yielding Full-length Purified Proteins
Published on: October 29, 2013
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Purification of GST-Tagged Proteins
Frank Schäfer1, Nicole Seip1, Barbara Maertens1
1QIAGEN GmbH, Research and Development, Qiagenstrasse 1, 40724 Hilden, Germany.
Methods in Enzymology
|June 23, 2015
Summary
This protocol details purifying glutathione S-transferase (GST) tagged proteins using glutathione affinity chromatography. This method enhances protein solubility and facilitates downstream applications like protein interaction studies.
Area of Science:
- Biochemistry
- Molecular Biology
- Protein Chemistry
Background:
- Glutathione S-transferase (GST) tags enhance recombinant protein solubility.
- GST-tagged proteins are crucial for functional characterization and interaction studies.
- Glutathione affinity purification is a standard method for isolating GST-tagged proteins.
Purpose of the Study:
- To describe a detailed protocol for purifying GST-tagged recombinant proteins.
- To provide a scalable method applicable to various expression systems.
- To offer guidance for optimizing purification from different cell types.
Main Methods:
- Utilizing glutathione affinity chromatography for purification.
- Employing a bind-wash-elute procedure for separation from cell lysate.
- Adapting the protocol for E. coli, insect, and mammalian expression systems.
Main Results:
- Successful purification of GST-tagged proteins from E. coli.
- Demonstration of the protocol's scalability for different expression levels and volumes.
- Guidance on modifying the protocol for diverse expression systems.
Conclusions:
- The described protocol provides an efficient and scalable method for GST-tagged protein purification.
- This technique is valuable for protein research, including interaction studies.
- The protocol is adaptable for various expression systems, enhancing its utility.
