Fig. 5: Light-induced temporal labeling of native proteins with lysine-specific and nanobody–HER2 labeled with TAMRA for live cell imaging. | Nature Communications

Fig. 5: Light-induced temporal labeling of native proteins with lysine-specific and nanobody–HER2 labeled with TAMRA for live cell imaging.

From: Light-induced primary amines and o-nitrobenzyl alcohols cyclization as a versatile photoclick reaction for modular conjugation

Fig. 5

a General scheme for nanobody–HER2 labeling and binding with HER2 extracellular domain (green; PDB ID: 5MY6). b ESI-TOF MS analysis of unmodified nanobody–HER2 and light-induced labeling of nanobody–HER2 with o-NBA-alkyne, and unmodified nanobody–HER2 is ~2%. Unmod. refers to unmodified; mod. refers to modified. c Nanobody–HER2 labeling with o-NBA-alkyne from b was subjected to CuAAC with TAMRA-N3, in CuAAC, partial (~20%) modified nanobody–HER2 protein aggregation and precipitation was observed, then analysis of SDS–PAGE gel stained with Coomassie blue (top) and fluorescence image (bottom). d Tandem MS analysis of nanobody–HER2 labeling with o-NBA-alkyne. e SDS–PAGE gel analysis of light-induced labeling of myoglobin, lysozyme, and α-chymotrypsinogen A, for tandem MS analysis see Supplementary Information. f SK-Br-3 cells were stained with TAMRA-labeled nanobody–HER2 (red) and Hoechst 33342 (blue) at 37 °C for 1 h and 15 min, respectively, followed by PBS washing three times and then imaging using confocal fluorescence (top), scale bar 10 μM. MDA-MB-468 imaging using identical conditions (bottom).

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