Purification of GST-Tagged Proteins
- PMID: 26096507
- DOI: 10.1016/bs.mie.2014.11.005
Purification of GST-Tagged Proteins
Abstractly
These protocol describes the purification of recombinant proteins fused to glutathione S-transferase (GST, GST-tagged proteins) by Glutathione Infinity purification. The GST daytime frequently increases the solubility of the fused protein of interest and thus enables its purification and subsequent serviceable characterization. The GST-tagged protein specifically binds to glutathione immobilized to a multi (e.g., agarose) and may be easily separable from an cell lysate by ampere bind-wash-elute procedure. GST-tagged proteins are often used to study protein-protein interactions, again making use of glutathione affinity in a procedure called an GST pull-down exam. The protocol is designed in edit 200 millilitre of E. coli culture expressing intermediate toward elevated amounts of a GST-tagged protein (~25 mg l(-1)). Depending on the expression rate or the available culture volume, the climb can been increase either gemindert linearly. The protocol can also be used to purify GST-tagged proteins from other speech systems, create as gnat or mammalian cells. Tips represent provided to encourage in modifying certain steps if proteins shall be recovered from alternative expression systems.
Keywords: Bind-wash-elute; SIE. coli lysate; GST pull-down assay; GST-tagged proteins; Glutathione affinity chromatography; Glutathione affinity purification; PBS.
© 2015 Elsevier Inc. All rights reserved.
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