Protein tag

About: Protein tag is a research topic. Over the lifetime, 553 publications have been published within this topic receiving 25944 citations.

Using Antibodies: A Laboratory Manual

Abstract: Preface Dedication Chapter 1: Antibody Structure And Function Chapter 2: Antibody-Antigen Interactions Chapter 3: Choosing Antibodies Chapter 4: Handling Antibodies Storing Antibodies Purifying Antibodies Labeling Antibodies Chapter 5: Staining Cells Major Constraints Choosing The Correct Antibody Protocols For Immunostaining Of Tissue Culture Cells Variations On The Immunostaining Procedure Chapter 6: Staining Tissues Major Constraints Choosing the Correct Antibody Immunostaining Tissues With Polyclonal Or Monoclonal Antibodies Protocols For Staining Tissue Sections Variations For Staining Yeast Variations For Staining Drosophila Protocols For Antigen Retrieval Chapter 7: Immunoprecipitation Major Constraints Choosing The Correct Antibody Immunoprecipitation Protocols Variations On The Immunoprecipitation Procedure Chapter 8: Immunoblotting Major Constraints Choosing The Correct Antibody Immunoblotting Protocols Variations In The Detection Methods Variations In The Immunoblotting Technique Chapter 9: Immunoaffinity Purification Major Constraints Choosing The Correct Antibody Immunoaffinity Purification With Polyclonal And Monoclonal Antibodies Protocols For Immunoaffinity Purification Variations On Immunoaffinity Purification Chapter 10: Tagging Proteins Major Constraints Why Tag Your Protein Tags For Protein Detection Tags For Protein Purification Choosing The Right Tag Deciding Where To Position The Tag How To Tag Your Protein Chapter 11: Epitope Mapping Determining The Structural Requirements Of An Epitope Choosing An Epitope-mapping Method Mapping By Competition Assay Mapping By Expression Of Gene Fragments Mapping Using Synthetic Peptides Synthesis Or Purchase Of The Peptide Set Alternate Approaches To Epitope Mapping Appendix I: Electrophoresis Appendix II: Protein Techniques Appendix III: General Information Appendix IV: Cautions Appendix V: Trademarks Appendix VI: Suppliers

HaloTag: A Novel Protein Labeling Technology for Cell Imaging and Protein Analysis

Abstract: We have designed a modular protein tagging system that allows different functionalities to be linked onto a single genetic fusion, either in solution, in living cells, or in chemically fixed cells. The protein tag (HaloTag) is a modified haloalkane dehalogenase designed to covalently bind to synthetic ligands (HaloTag ligands). The synthetic ligands comprise a chloroalkane linker attached to a variety of useful molecules, such as fluorescent dyes, affinity handles, or solid surfaces. Covalent bond formation between the protein tag and the chloroalkane linker is highly specific, occurs rapidly under physiological conditions, and is essentially irreversible. We demonstrate the utility of this system for cellular imaging and protein immobilization by analyzing multiple molecular processes associated with NF-κB-mediated cellular physiology, including imaging of subcellular protein translocation and capture of protein−protein and protein−DNA complexes.

A general method for the covalent labeling of fusion proteins with small molecules in vivo

Abstract: Characterizing the movement, interactions, and chemical microenvironment of a protein inside the living cell is crucial to a detailed understanding of its function. Most strategies aimed at realizing this objective are based on genetically fusing the protein of interest to a reporter protein that monitors changes in the environment of the coupled protein. Examples include fusions with fluorescent proteins, the yeast two-hybrid system, and split ubiquitin. However, these techniques have various limitations, and considerable effort is being devoted to specific labeling of proteins in vivo with small synthetic molecules capable of probing and modulating their function. These approaches are currently based on the noncovalent binding of a small molecule to a protein, the formation of stable complexes between biarsenical compounds and peptides containing cysteines, or the use of biotin acceptor domains. Here we describe a general method for the covalent labeling of fusion proteins in vivo that complements existing methods for noncovalent labeling of proteins and that may open up new ways of studying proteins in living cells.

Overview of tag protein fusions: from molecular and biochemical fundamentals to commercial systems.

Abstract: In response to the rapidly growing field of proteomics, the use of recombinant proteins has increased greatly in recent years. Recombinant hybrids containing a polypeptide fusion partner, termed affinity tag, to facilitate the purification of the target polypeptides are widely used. Many different proteins, domains, or peptides can be fused with the target protein. The advantages of using fusion proteins to facilitate purification and detection of recombinant proteins are well-recognized. Nevertheless, it is difficult to choose the right purification system for a specific protein of interest. This review gives an overview of the most frequently used and interesting systems: Arg-tag, calmodulin-binding peptide, cellulose-binding domain, DsbA, c-myc-tag, glutathione S-transferase, FLAG-tag, HAT-tag, His-tag, maltose-binding protein, NusA, S-tag, SBP-tag, Strep-tag, and thioredoxin.