Titlebook: Site-Specific Protein Labeling; Methods and Protocol Arnaud Gautier,Marlon J. Hinner Book 2015 Springer Science+Business Media New York 201

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HaloTag Technology for Specific and Covalent Labeling of Fusion Proteins,he last few decades many varieties of fluorescent proteins have been generated, each bringing new capability to research. However, taking full advantage of standard fluorescent proteins with advanced and differential features requires significant effort on the part of the researcher. This approach n

Metamorphosis

Ligation of Synthetic Peptides to Proteins Using Semisynthetic Protein ,-Splicing,ns. In particular, semisynthetic protein .-splicing with one intein fragment short enough to be accessible by solid-phase peptide synthesis can be used to transfer a short peptide segment with the desired synthetic moiety to the protein of interest. In this chapter, we provide detailed protocols for

manifestation

Chemical-Tag Labeling of Proteins Using Fully Recombinant Split Inteins,f any chemical synthesis to be performed. In a two-step protocol, a very short tag fused to a split intein auxiliary protein is first labeled in a bioconjugation reaction with a synthetic moiety either at its N-terminus (amine-tag) or at the side chain of an unnatural amino acid (click-tag). The lab

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Oratory

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Site-Specific Labeling of Proteins via Sortase: Protocols for the Molecular Biologist,hods, while versatile with respect to the types of moieties that can be attached, suffer from lack of specificity, often targeting multiple positions within a protein. Here we describe protocols for the chemoenzymatic labeling of proteins at the C-terminus using the bacterial transpeptidase, sortase

Malleable

BONCAT: Metabolic Labeling, Click Chemistry, and Affinity Purification of Newly Synthesized Proteomological processes but cannot be combined with modern high-throughput mass spectrometry analysis. This chapter describes the unbiased identification of a whole de novo synthesized proteome of cultured cells or of a translationally active subcellular fraction of the mammalian brain. This technique re

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Genetic Encoding of Unnatural Amino Acids for Labeling Proteins, proteins with any probe. Here we describe the genetic encoding of dienophile-bearing unnatural amino acids into proteins expressed in . and mammalian cells using the pyrrolysyl-tRNA synthetase/tRNA. pair and its variants. We describe the rapid fluorogenic labeling of proteins containing these unnat

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infatuation