Protein purification tags

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== amino acid tags ==
== amino acid tags ==
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# His tag - several histidines
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=== Principle ===
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A sequence of amino acids is added to the protein and enables the recovery of the labelled protein by its unique affinity. Usually its easiest to add the stretch of amino acids to the ends of the protein to ensure its accessibility and not to disturb the protein folding.
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===Examples===
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=== His tag ===
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:6+ histidine (His) residues
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:recovered by affinity to nickel or cobalt column
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<small>
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:[[wikipedia:Polyhistidine-tag|more on the ''His tag'' in the Wikipedia]]
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</small>
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=== FLAG tag ===
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:N-DYKDDDDK-C and others
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:recovered with specific antibody
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<small>
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:[[wikipedia:FLAG-tag|- more on ''FLAG-tag'' in the Wikipedia]]
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:- protocol for purification of His-tagged protein, Kafatos lab: [[Kafatos:His-tagged Protein Purification (denaturing)]]
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:- protocol for purification of His-tagged protein, Wittrup lab: [[Wittrup:His-Tag Proteins/Nickel Column Purification (suppliers, troubleshooting, etc.)]]
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</small>
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=== HA tag ===
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=== GST tag ===
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:fusion with the small glutathione-S-transferase (GST; 26 kDa)
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:recovery by affinity to substrate glutathione bound to a column, e.g. glutathione sepharose
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<small>
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:- GST system of GE Healthcare [http://www6.gelifesciences.com/APTRIX/upp01077.nsf/Content/Products?OpenDocument&parentid=366157&moduleid=164393&zone=Proteomics]
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</small>
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=== myc tag ===
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:
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<small>
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:-
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</small>
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=== biotin - streptavidin ===
== chemical tags ==
== chemical tags ==
== links ==
== links ==
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=== Wikipedia ===
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* [[wikipedia:YY|XX]]
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* [[wikipedia:Protein tag|Protein tag]]
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* [[wikipedia:Protein_purification#Purification_of_a_tagged_protein|Purification of a tagged protein]]
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* Wikipedia: [[wikipedia:Polyhistidine-tag|His tag]]
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=== OWW ===
* example of a protocol for protein purification from bacteria: [[Protein Expression and Purification]]
* example of a protocol for protein purification from bacteria: [[Protein Expression and Purification]]
* list but no content of purification methods from Berglund lab: [[Berglund:Protein Purification: Various Methods]]
* list but no content of purification methods from Berglund lab: [[Berglund:Protein Purification: Various Methods]]
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* protocol for purification of His-tagged protein, Kafatos lab: [[Kafatos:His-tagged Protein Purification (denaturing)]]
 

Revision as of 16:57, 3 March 2007

Overview of tags added to protein to facilitate their purification

Contents

amino acid tags

Principle

A sequence of amino acids is added to the protein and enables the recovery of the labelled protein by its unique affinity. Usually its easiest to add the stretch of amino acids to the ends of the protein to ensure its accessibility and not to disturb the protein folding.

Examples

His tag

6+ histidine (His) residues
recovered by affinity to nickel or cobalt column

more on the His tag in the Wikipedia

FLAG tag

N-DYKDDDDK-C and others
recovered with specific antibody

- more on FLAG-tag in the Wikipedia
- protocol for purification of His-tagged protein, Kafatos lab: Kafatos:His-tagged Protein Purification (denaturing)
- protocol for purification of His-tagged protein, Wittrup lab: Wittrup:His-Tag Proteins/Nickel Column Purification (suppliers, troubleshooting, etc.)

HA tag

GST tag

fusion with the small glutathione-S-transferase (GST; 26 kDa)
recovery by affinity to substrate glutathione bound to a column, e.g. glutathione sepharose

- GST system of GE Healthcare [1]


myc tag

-

biotin - streptavidin

chemical tags

links

Wikipedia

OWW

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