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Protocols
- Two Hybrid System Transformation (Gietz Lab)
This Protocol is for 2 Hybrid system screens
http://www.umanitoba.ca/faculties/medicine/biochem...
Added: Tue May 14 2002, Hits: 823, Reviews: 0
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Co-Immunoprecipitation Analysis of Protein-Protein Interaction
(Morrisey Lab, University of Pennsylvania)
This protocol uses total cell extracts to analyze putative protein-protein interactions in eukaryotic cells. One can also use nuclear extracts as a source for this protocol.
http://www.uphs.upenn.edu/mcrc/morrisey/coip
Added: Mon Oct 14 2002, Hits: 20116, Reviews: 0
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GST Fusion Protein Purification from Yeast
(Dohlman Lab)
http://www.med.unc.edu/~hdohlman/gst.html
Added: Tue May 14 2002, Hits: 1228, Reviews: 0
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Performing a hunt by interaction mating
(Finley Lab, Wayne State University, School of Medicine)
In this protocol one strain is transformed with library DNA and the transformants are collected and frozen in aliquots. For each interactor hunt, an aliquot of this frozen "pretransformed library strain" is thawed and mixed with an aliquot of a bait strain transformed with the bait expression plasmid. Overnight incubation of the mixture on a YPD plate results in mating - i.e., cells of one strain fuse with cells of the other strain to form diploids. The diploids are then exposed to galactose to induce expression of the library encoded proteins, and interactors are selected in the same manner as in the standard hunt protocols.
http://proteome.wayne.edu/Matinghunt.htm
Added: Tue May 14 2002, Hits: 98, Reviews: 0
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Protein Interaction Analysis Using an IASYS Biosensor
(Kitto Lab, The University of Texas at Austin)
Immobilization of ligands on cuvette surfaces and measure the interactions of ligand which is immobilized on the cuvette and the ligate which is added to the immobilized cuvette.
http://kitto.cm.utexas.edu/research/Kittolabpage/P...
Added: Tue Oct 08 2002, Hits: 918, Reviews: 0
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Cached - Purification of GST Fused Proteins (Contributed by Chia Jin Ngee)
Many people have vented out frustration over insoluble GST-fused proteins. This is a protocol for enzymatically active soluble GST-fused proteins. All GST-fused proteins are rendered soluble with this technique though enzyme activitiy can range from 30-90%.
Added: Tue May 14 2002, Reviews: 0
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Purification of GST Fused Proteins
(Bowtell Lab Manual, Peter MacCallum Cancer Institute)
http://www.pmci.unimelb.edu.au/core_facilities/man...
Added: Tue May 14 2002, Hits: 228, Reviews: 0
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Purification of His-tagged proteins from Sf9 cells under native conditions
(Brent Graveley)
http://penguin.uchc.edu/~intron/Protocols/Sf9%20Hi...
Added: Tue May 14 2002, Hits: 502, Reviews: 0
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Purification of MBP (maLTose-Binding Proteins) Fused Proteins
(Bowtell Lab Manual, Peter MacCallum Cancer Institute)
http://www.pmci.unimelb.edu.au/core_facilities/man...
Added: Tue May 14 2002, Hits: 81, Reviews: 0
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Quantitative immunoprecipitation of GFP-fusion proteins using the GFP-Trap
(Epigenome Network of Excellence)
This protocol, using the versatile GTP-Trap, can be used to perform immunoprecipitation of fluorescent fusion proteins from cell extracts to identify and map protein-protein interactions as exemplarily shown by the dimerization of Dnmt1
http://www.epigenome-noe.net/researchtools/protoco...
Added: Wed May 27 2009, Hits: 1431, Reviews: 0
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Yeast Two-Hybrid System
(Finley Lab, Wayne State University, School of Medicine)
This is one of the most comprehensive and detailed guide to yeast two-hybrid system technique with introduction to its background. The following procedures are described: mating assay - small scale for tens of different bait or prey proteins, collecting bait (and prey) strains, large scale for hundreds of different bait or prey strains, interaction mating assay with other yeast two-hybrid systems, recording results, interpreting interaction data, qualitative interpretation and more...
http://proteome.wayne.edu/YTHnetworks.html
Added: Tue May 14 2002, Hits: 1150, Reviews: 0
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Yeast Two-Hybrid System
(Finley Lab, Wayne State University, School of Medicine)
The updated version for scaling up the these techniques to facilitate characterization of large networks of interacting proteins. It is also useful for individual hunts.
http://proteome.wayne.edu/Update.html
Added: Tue May 14 2002, Hits: 825, Reviews: 0
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Yeast Two-Hybrid System: part A: screen preparation
(Technical Tips Online)
(To view this page, free registration is needed)
The two-hybrid system (THS) is a powerful technique for identifying new proteins involved in specific biological processes. It allows for the rapid isolation of the gene that codes for a protein that interacts with a specific protein of interest.
http://tto.biomednet.com/cgi-bin/tto/pr/pg_art.cgi...
Added: Tue May 14 2002, Hits: 856, Reviews: 0
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