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Full Description
Protein Affinity Tags: Methods and Protocols provides researchers with the necessary information, tools, and strategy required for proper inquiry into a given protein's function and structure. Today's researchers can easily alter the amino acid sequence of any given protein, a powerful technology allowing for the precise engineering of specific proteins. Protein affinity tagging employs the use of known protein binding interactions in order to "fish out" a protein-of-interest or, with more advanced tags, a protein complex. This technology paired with recent advancements in DNA sequencing technology promises a future that is awash with novel genomic information - information that will not only expand our knowledge of the processes that govern life, but also fuel innovation that will undoubtedly conquer many of today's most salient health and environmental challenges. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.
Authoritative and easily accessible, Protein Affinity Tags: Methods and Protocols will be an invaluable resource to those seeking to employ protein affinity tags to study their biological system of interest.
Contents
1D4 - A Versatile Epitope Tag for the Purification and Characterization of Expressed Membrane and Soluble Proteins.- Affinity Purification of Heme-Tagged Proteins.- Purification of a Recombinant Protein with Cellulose-Binding Module 3 as the Affinity Tag.- Purification of E. coli Proteins Using a Self-Cleaving Chitin-Binding Affinity Tag.- Simplified Protein Purification Using an Autoprocessing, Inducible Enzyme Tag.- SUMO as a Solubility Tag and in vivo Cleavage of SUMO Fusion Proteins with Ulp1.- Rescuing Aggregation-Prone Proteins in Escherichia coli with a Dual His6-MBP Tag.- Expression, Purification, and Immobilization of Recombinant Tamavidin 2 Fusion Proteins.- Use of Tandem Affinity Chromatography for Purification of Cannabinoid Receptor CB2.- Detection of Protein-Protein Interactions Using Tandem Affinity Purification.- An Improved in vivo Biotinylation Strategy Combined with FLAG and Antibody Based Approaches for Affinity Purification of Protein Complexes in Mouse Embryonic Stem Cells.- Purification of Recombinant Proteins with a Multifunctional GFP Tag.- Targeted Purification of SnAvi-Tagged Proteins.- An Efficient Fluorescent Protein-Based Multifunctional Affinity Purification Approach in Mammalian Cells.- Bimolecular Affinity Purification - A Variation of TAP with Multiple Applications.
