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Protein Affinity Tags

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Cover of 'Protein Affinity Tags'

Table of Contents

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    Book Overview
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    Chapter 1 1D4: A Versatile Epitope Tag for the Purification and Characterization of Expressed Membrane and Soluble Proteins
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    Chapter 2 Affinity Purification of Heme-Tagged Proteins
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    Chapter 3 Purification of a recombinant protein with cellulose-binding module 3 as the affinity tag.
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    Chapter 4 Purification of E. coli Proteins Using a Self-Cleaving Chitin-Binding Affinity Tag
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    Chapter 5 Simplified Protein Purification Using an Autoprocessing, Inducible Enzyme Tag
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    Chapter 6 SUMO as a Solubility Tag and In Vivo Cleavage of SUMO Fusion Proteins with Ulp1.
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    Chapter 7 Rescuing Aggregation-Prone Proteins in Escherichia coli with a Dual His6-MBP Tag.
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    Chapter 8 Expression, Purification, and Immobilization of Recombinant Tamavidin 2 Fusion Proteins
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    Chapter 9 Use of Tandem Affinity Chromatography for Purification of Cannabinoid Receptor CB 2
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    Chapter 10 Detection of protein-protein interactions using tandem affinity purification.
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    Chapter 11 An Improved In Vivo Biotinylation Strategy Combined with FLAG and Antibody Based Approaches for Affinity Purification of Protein Complexes in Mouse Embryonic Stem Cells
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    Chapter 12 Purification of Recombinant Proteins with a Multifunctional GFP Tag
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    Chapter 13 Targeted Purification of SnAvi-Tagged Proteins
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    Chapter 14 An efficient fluorescent protein-based multifunctional affinity purification approach in Mammalian cells.
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    Chapter 15 Bimolecular Affinity Purification: A Variation of TAP with Multiple Applications
Attention for Chapter 10: Detection of protein-protein interactions using tandem affinity purification.
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Chapter title
Detection of protein-protein interactions using tandem affinity purification.
Chapter number 10
Book title
Protein Affinity Tags
Published in
Methods in molecular biology, June 2014
DOI 10.1007/978-1-4939-1034-2_10
Pubmed ID
Book ISBNs
978-1-4939-1033-5, 978-1-4939-1034-2
Authors

Goodfellow I, Bailey D, Ian Goodfellow, Dalan Bailey

Abstract

Tandem affinity purification (TAP) is an invaluable technique for identifying interaction partners for an affinity tagged bait protein. The approach relies on the fusion of dual tags to the bait before separate rounds of affinity purification and precipitation. Frequently two specific elution steps are also performed to increase the specificity of the overall technique. In the method detailed here, the two tags used are protein G and a short streptavidin binding peptide; however, many variations can be employed. In our example the tags are separated by a cleavable tobacco etch virus protease target sequence, allowing for specific elution after the first round of affinity purification. Proteins isolated after the final elution step in this process are concentrated before being identified by mass spectrometry. The use of dual affinity tags and specific elution in this technique dramatically increases both the specificity and stringency of the pull-downs, ensuring a low level of background nonspecific interactions.

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Mendeley readers

The data shown below were compiled from readership statistics for 10 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 10 100%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 3 30%
Student > Bachelor 3 30%
Student > Master 2 20%
Other 1 10%
Researcher 1 10%
Other 0 0%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 5 50%
Agricultural and Biological Sciences 2 20%
Chemistry 1 10%
Neuroscience 1 10%
Unknown 1 10%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 13 February 2015.
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#20,259,845
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Outputs from Methods in molecular biology
#9,896
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#192,913
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Outputs of similar age from Methods in molecular biology
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