Protein purification tags: Difference between revisions
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== amino acid tags == | == 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 | |||
<small> | |||
:[[wikipedia:Polyhistidine-tag|more on the ''His tag'' in the Wikipedia]] | |||
</small> | |||
=== FLAG tag === | |||
:N-DYKDDDDK-C and others | |||
:recovered with specific antibody | |||
<small> | |||
:[[wikipedia:FLAG-tag|- 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.)]] | |||
</small> | |||
=== 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 | |||
<small> | |||
:- GST system of GE Healthcare [http://www6.gelifesciences.com/APTRIX/upp01077.nsf/Content/Products?OpenDocument&parentid=366157&moduleid=164393&zone=Proteomics] | |||
</small> | |||
=== myc tag === | |||
: | |||
<small> | |||
:- | |||
</small> | |||
=== biotin - streptavidin === | |||
== chemical tags == | == chemical tags == | ||
== links == | == links == | ||
=== Wikipedia === | |||
* [[wikipedia:YY|XX]] | |||
* [[wikipedia:Protein tag|Protein tag]] | |||
* [[wikipedia:Protein_purification#Purification_of_a_tagged_protein|Purification of a tagged protein]] | |||
=== 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]] | ||
Revision as of 14:57, 3 March 2007
Overview of tags added to protein to facilitate their purification
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
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
- 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
