Table 1.

Major puromycin derivatives.

Compound	Description	Uses
Nucleoside substitutions
5′ Fluorophore-dC-puromycin	Cy3, Cy5, or fluorescein attached via deoxycytidine linker	Imaging protein synthesis in cultured cells [23]; In vitro C-terminal labeling of full-length proteins [24]
5′ Biotin-dC-puromycin	Biotin attached via deoxycytidine linker	Labeling newly synthesized proteins under cell free conditions for subsequent affinity purification and proteomic analysis [25], [26]
5′ alkyne/azide puromycin (5Y/5Z/5N/5T)	Alkyne/azide substitution of the 5′ hydroxyl group	Labeling newly synthesized proteins in cultured cells for visualization or affinity purification using click chemistry cycloaddition of fluorophore/biotin [19]
Amino acid substitutions
O-propargyl-puromycin (OPP)	Alkyne substitution of the O-methyl-phenyl ring	Labeling newly synthesized proteins in cultured cells, tissues and whole animals for visualization or affinity purification using click chemistry cycloaddition of fluorophore/biotin [8], [27]
Photocleavable N-blocked (NVOC/DEACM) puromycin	Photocleavable group attached to the free amino terminus	Labeling newly synthesized proteins in cultured cells with improved spatiotemporal resolution using laser excitation [17], [18]
Enzyme labile N-blocked puromycin (PhAc-puro)	Enzyme labile phenylacetyl group attached to the free amino terminus	Selective labeling of newly synthesized proteins in cultured cells engineered to express an E. coli enzyme [28]
N-blocked biotin puromycin (3P)	Biotin attached to the free amino terminus via double Jeffamine linker	Affinity purification of ribosomes under cell free conditions (without puromycylation) [16]