Protein purification tags - Revision history

Christopher C Vanlang at 01:55, 30 December 2011

Christopher C Vanlang — Fri, 30 Dec 2011 01:55:31 GMT

←Older revision		Revision as of 01:55, 30 December 2011
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	* [http://www.the-scientist.com/article/display/12881/ article in ''The Scientist'' on protein purification with tags, 2002]		* [http://www.the-scientist.com/article/display/12881/ article in ''The Scientist'' on protein purification with tags, 2002]

-	[[Category:Material]] [[Category:Miscellaneous]]	+	[[Category:Material]] [[Category:Miscellaneous]][[Category:Purification]]

Torsten Waldminghaus at 21:00, 25 February 2009

Torsten Waldminghaus — Wed, 25 Feb 2009 21:00:27 GMT

←Older revision		Revision as of 21:00, 25 February 2009
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	=== other ===		=== other ===
	* [http://www.the-scientist.com/article/display/12881/ article in ''The Scientist'' on protein purification with tags, 2002]		* [http://www.the-scientist.com/article/display/12881/ article in ''The Scientist'' on protein purification with tags, 2002]
		+
		+	[[Category:Material]] [[Category:Miscellaneous]]

Jakob Suckale: suggestions for improvement

Jakob Suckale — Sat, 03 Mar 2007 23:09:36 GMT

suggestions for improvement

←Older revision		Revision as of 23:09, 3 March 2007
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	'''Overview of tags added to protein to facilitate their purification'''		'''Overview of tags added to protein to facilitate their purification'''

-	== ~~Amino~~ acid tags ==	+	We are missing info on TAP-tag and evalution/comparison of the techniques. [[User:Jasu\|Jasu]] 18:09, 3 March 2007 (EST)
		+
		+	== amino acid tags ==

	A stretch 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 tag to either end of the protein to ensure its accessibility and not to disturb the protein folding.		A stretch 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 tag to either end of the protein to ensure its accessibility and not to disturb the protein folding.
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	</small>		</small>

-	== ~~Protein~~ tags ==	+	== protein tags ==

	Rather than adding only a few amino acids a whole protein is fused to the protein to be purified or detected. The affinity of the attached protein enables the recovery of the artificial fusion protein. As for the peptides, the protein tag is added to either end of the target protein.		Rather than adding only a few amino acids a whole protein is fused to the protein to be purified or detected. The affinity of the attached protein enables the recovery of the artificial fusion protein. As for the peptides, the protein tag is added to either end of the target protein.

Jakob Suckale: * page creation * finished - up to you to extend & improve ;-)

Jakob Suckale — Sat, 03 Mar 2007 23:07:06 GMT

*** page creation *** finished - up to you to extend & improve ;-)

←Older revision		Revision as of 23:07, 3 March 2007
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	A stretch 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 tag to either end of the protein to ensure its accessibility and not to disturb the protein folding.		A stretch 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 tag to either end of the protein to ensure its accessibility and not to disturb the protein folding.

-	=== His tag ===	+	=== His tag / IMAC ===

	:6+ histidine (His) residues		:6+ histidine (His) residues
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	</small>		</small>

-	=== HA tag ===	+	=== HA tag===
	:fusion with an epitope derived from the Influenza protein haemagglutinin (HA): e.g. N-YPYDVP-C		:fusion with an epitope derived from the Influenza protein haemagglutinin (HA): e.g. N-YPYDVP-C
	:recovery with an HA antibody		:recovery with an HA antibody
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	:- [[wikipedia:Glutathione S-transferase\|Wikipedia entry on GST]]		:- [[wikipedia:Glutathione S-transferase\|Wikipedia entry on GST]]
	</small>		</small>
		+
		+	== chemical tags ==

	=== biotin - streptavidin ===		=== biotin - streptavidin ===
-	:	+	:linking amino acids, often lysines, to cofactor and vitamin biotin
		+	:purification by tight binding to streptavidin-agarose or -beads
	<small>		<small>
-	:- [[wikipedia:XX\|YY]]	+	:- [[wikipedia:Biotin\|Wikipedia entry biotin]]
-	:-	+	:- [[wikipedia:Streptavidin\|Wikipedia entry streptavidin]] and [[wikipedia:Avidin\|avidin]]
		+	:- [http://www.iba-go.com/iba_us/prottools/prot_p_bur.html Biotin purification reagents from IBA]
	</small>		</small>
-
-	~~== chemical tags ==~~

	== links ==		== links ==
	=== Wikipedia ===		=== Wikipedia ===
-	* [[wikipedia:YY\|XX]]
-
	* [[wikipedia:Protein tag\|Protein tag]]		* [[wikipedia:Protein tag\|Protein tag]]
	* [[wikipedia:Protein_purification#Purification_of_a_tagged_protein\|Purification of a tagged protein]]		* [[wikipedia:Protein_purification#Purification_of_a_tagged_protein\|Purification of a tagged protein]]

	=== OWW ===		=== OWW ===
-	* example of a protocol for protein purification from bacteria~~: [[Protein Expression and Purification~~]]	+	* [[Protein Expression and Purification\|example of a protocol for protein purification from bacteria]]
-	* list but no content of purification methods from Berglund lab: [~~[Berglund~~:~~Protein Purification: Various Methods]~~]	+	* [[Berglund:Protein Purification: Various Methods\|list but no content of purification methods from Berglund lab]]
		+
		+	=== other ===
		+	* [http://www.the-scientist.com/article/display/12881/ article in ''The Scientist'' on protein purification with tags, 2002]

Jakob Suckale: * page creation * unfinished save 3

Jakob Suckale — Sat, 03 Mar 2007 22:38:53 GMT

*** page creation *** unfinished save 3

←Older revision		Revision as of 22:38, 3 March 2007
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	'''Overview of tags added to protein to facilitate their purification'''		'''Overview of tags added to protein to facilitate their purification'''

-	== ~~amino~~ acid tags ==	+	== Amino acid tags ==

-	~~=== Principle ===~~	+	A stretch 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 tag to either end of the protein to ensure its accessibility and not to disturb the protein folding.

-	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 ===		=== His tag ===

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	:recovered by affinity to nickel or cobalt column		:recovered by affinity to nickel or cobalt column
	<small>		<small>
-	:[[wikipedia:Polyhistidine-tag\|more on the ''His tag'' in the Wikipedia]]	+	:- [[wikipedia:Polyhistidine-tag\|more on the ''His tag'' in the Wikipedia]]
	</small>		</small>

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	:recovered with specific antibody		:recovered with specific antibody
	<small>		<small>
-	:[[wikipedia:FLAG-tag\|- more on ''FLAG-tag'' in the Wikipedia]]	+	:- [[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)]]	+	:- [[Kafatos:His-tagged Protein Purification (denaturing)\|protocol for purification of His-tagged protein, Kafatos lab]]
-	:- protocol for purification of His-tagged protein, ~~Wittrup~~ lab: [[Wittrup:His-Tag Proteins/Nickel Column Purification (suppliers, troubleshooting, etc.)]]	+	:- [[Wittrup:His-Tag Proteins/Nickel Column Purification (suppliers, troubleshooting, etc.)\|protocol for purification of His-tagged protein, Wittrup lab]]
	</small>		</small>

	=== HA tag ===		=== HA tag ===
		+	:fusion with an epitope derived from the Influenza protein haemagglutinin (HA): e.g. N-YPYDVP-C
		+	:recovery with an HA antibody
		+	<small>
		+	:- [[wikipedia:Hemagglutinin\|Wikipedia entry on haemagglutinin]]
		+	:- [http://www.clontech.com/products/detail.asp?tabno=2&catalog_id=631207&page=all Clontech: HA tag]
		+	:- [http://www.roche-applied-science.com/sis/proteinexpression/docs/manual_ha.pdf Roche HA tag vector (PDF)]
		+	</small>
		+
		+	=== MYC tag ===
		+	:fusion with an epitope derived from the human proto-oncoprotein MYC: e.g. N-ILKKATAYIL-C, N-EQKLISEEDL-C
		+	:recovery with an MYC antibody
		+	<small>
		+	:- [[wikipedia:MYC\|Wikipedia entry on Myc oncogene]]
		+	:- [http://www.miltenyibiotec.com/download/mm366/DS130-092-085.pdf MACS-MYC vector from Miltenyi Biotec (PDF)]
		+	:- [http://www.stratagene.com/products/displayProduct.aspx?pid=242 combined MYC/FLAG tag vector from Stratagene]
		+	</small>
		+
		+	== Protein tags ==
		+
		+	Rather than adding only a few amino acids a whole protein is fused to the protein to be purified or detected. The affinity of the attached protein enables the recovery of the artificial fusion protein. As for the peptides, the protein tag is added to either end of the target protein.

	=== GST tag ===		=== GST tag ===
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	:recovery by affinity to substrate glutathione bound to a column, e.g. glutathione sepharose		:recovery by affinity to substrate glutathione bound to a column, e.g. glutathione sepharose
	<small>		<small>
-	:- ~~GST system of GE Healthcare~~ [http://www6.gelifesciences.com/APTRIX/upp01077.nsf/Content/Products?OpenDocument&parentid=366157&moduleid=164393&zone=Proteomics]	+	:- [http://www6.gelifesciences.com/APTRIX/upp01077.nsf/Content/Products?OpenDocument&parentid=366157&moduleid=164393&zone=Proteomics GST cloning system of GE Healthcare]
		+	:- [[wikipedia:Glutathione S-transferase\|Wikipedia entry on GST]]
	</small>		</small>

-		+	=== biotin - streptavidin ===
-	=== ~~myc tag~~ ===	+
	:		:
	<small>		<small>
-	:-	+	:- [[wikipedia:XX\|YY]]
		+	:-
	</small>		</small>
-
-	~~=== biotin - streptavidin ===~~

	== chemical tags ==		== chemical tags ==

Jakob Suckale: * page creation * unfinished save 2

Jakob Suckale — Sat, 03 Mar 2007 21:57:11 GMT

*** page creation *** unfinished save 2

←Older revision		Revision as of 21:57, 3 March 2007
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	== amino acid tags ==		== amino acid tags ==

-	# His tag - ~~several histidines~~	+	=== 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]]

-	* Wikipedia: [[wikipedia:Polyhistidine-tag\|His tag]]	+	=== 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]]
-	* protocol for purification of His-tagged protein, Kafatos lab: [[Kafatos:His-tagged Protein Purification (denaturing)]]

Jakob Suckale: * page creation * unfinished save 1

Jakob Suckale — Sat, 03 Mar 2007 21:24:28 GMT

*** page creation *** unfinished save 1

New page

'''Overview of tags added to protein to facilitate their purification'''

== amino acid tags ==

# His tag - several histidines

== chemical tags ==

== links ==

* Wikipedia: [[wikipedia:Polyhistidine-tag|His tag]]
* 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]]
* protocol for purification of His-tagged protein, Kafatos lab: [[Kafatos:His-tagged Protein Purification (denaturing)]]

Protein purification tags - Revision history

Christopher C Vanlang at 01:55, 30 December 2011

Torsten Waldminghaus at 21:00, 25 February 2009

Jakob Suckale: suggestions for improvement

Jakob Suckale: *** page creation *** finished - up to you to extend & improve ;-)

Jakob Suckale: *** page creation *** unfinished save 3

Jakob Suckale: *** page creation *** unfinished save 2

Jakob Suckale: *** page creation *** unfinished save 1

Jakob Suckale: * page creation * finished - up to you to extend & improve ;-)

Jakob Suckale: * page creation * unfinished save 3

Jakob Suckale: * page creation * unfinished save 2

Jakob Suckale: * page creation * unfinished save 1