Fluorogenic Tetrazine-Siliconrhodamine Probe for the Labeling of Noncanonical Amino Acid Tagged Proteins

Tetrazine-bearing fluorescent labels enable site-specific tagging of proteins that are genetically manipulated with dienophile modified noncanonical amino acids. The inverse electron demand Diels-Alder reaction between the tetrazine and the dienophile fulfills the criteria of bioorthogonality allowi...

Full description

Saved in:
Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.) Vol. 1728; p. 337
Main Authors Kozma, Eszter, Paci, Giulia, Estrada Girona, Gemma, Lemke, Edward A, Kele, Péter
Format Journal Article
LanguageEnglish
Published United States 2018
Subjects
Amino Acids - chemistry
Amino Acids - genetics
Animals
Cercopithecus aethiops
COS Cells
Fluorescent Dyes - chemical synthesis
Fluorescent Dyes - chemistry
Humans
Microscopy, Fluorescence - methods
Proteins - chemistry
Proteins - genetics
Rhodamines - chemistry
Silicon - chemistry
Staining and Labeling
Transfection
NIR emission
Super-resolution microscopy
Fluorogenicity
Bioorthogonality
Tetrazine
Online AccessGet more information

Cover

More Information
Summary:Tetrazine-bearing fluorescent labels enable site-specific tagging of proteins that are genetically manipulated with dienophile modified noncanonical amino acids. The inverse electron demand Diels-Alder reaction between the tetrazine and the dienophile fulfills the criteria of bioorthogonality allowing fluorescent labeling schemes of live cells. Here, we describe the detailed synthetic and labeling protocols of a near infrared emitting siliconrhodamine-tetrazine probe suitable for super-resolution imaging of residue-specifically engineered proteins in mammalian cells.
ISSN:1940-6029
DOI:10.1007/978-1-4939-7574-7_22