2013
DOI: 10.1002/0471140864.ps0909s73
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Overview of Affinity Tags for Protein Purification

Abstract: Addition of an affinity tag is a useful method for differentiating recombinant proteins expressed in bacterial and eukaryotic expression systems from the background of total cellular proteins, and for detecting protein‐protein interactions. This overview describes the historical basis for the development of affinity tags, affinity tags that are commonly used today, how to choose an appropriate affinity tag for a particular purpose, and several recently developed affinity tag technologies that may prove useful … Show more

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Cited by 248 publications
(166 citation statements)
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“…Therefore, they must be purified so as achieve conveniently high concentration and purity levels required for stable storage and characterization or for efficient use in biological systems or in catalysis. The identification of peptide tags for single-step affinity-based purification, upon end terminal fusion, has provided an advantageous tool that represents a faster, easy and cost-effective alternative to multiple-step chromatographic separations [1,2]. Protein tagging allows the purification of virtually any protein without previous knowledge of its properties or those of its expected contaminants, since the capturing event relies on tag's rather than on protein's features.…”
Section: Affinity Protein Purificationmentioning
confidence: 99%
“…Therefore, they must be purified so as achieve conveniently high concentration and purity levels required for stable storage and characterization or for efficient use in biological systems or in catalysis. The identification of peptide tags for single-step affinity-based purification, upon end terminal fusion, has provided an advantageous tool that represents a faster, easy and cost-effective alternative to multiple-step chromatographic separations [1,2]. Protein tagging allows the purification of virtually any protein without previous knowledge of its properties or those of its expected contaminants, since the capturing event relies on tag's rather than on protein's features.…”
Section: Affinity Protein Purificationmentioning
confidence: 99%
“…Even when satisfactory antibodies are generated, downstream experimental procedures need to be optimized for each new antibody. The second method is to express the protein of interest with a well-characterized epitope such as V5, FLAG or Myc3. This enables researchers to use commercially available, validated reagents and protocols for downstream applications.…”
mentioning
confidence: 99%
“…If the introduced protein is to be visualized by western blot, microscopy, or used for affinity purification, the protein should have a tag 35 for affinity purification or an available antibody for detection. However, for unknown reasons, some interactions known from the literature or other biochemical methods (data not shown) were not able to be recapitulated after electroporation.…”
Section: Discussionmentioning
confidence: 99%