Synthesis and Validation of Cyanine-Based Dyes for DIGE
The application of difference gel electrophoresis (DIGE), in particular its most common “minimal labeling” variety, utilizes N-hydroxysuccinimide esters of Cy2, Cy3, and Cy5 dyes, which are commercially available. We describe methods for the efficient synthesis of all three dyes from relatively inexpensive and commercially available precursors in only a few steps and with relatively high yields. In model DIGE experiments, the newly synthesized dyes proved to be indistinguishable from commercially available ones and have been shown to be stable for years while stored under argon as dry solids or after being dissolved in N,N-dimethylformamide.
Key wordsDIGE Difference gel electrophoresis Benzoxazolium Cy2 Propyl Cy3 Ethyl Cy5 Cy dye synthesis
Wan-Joong Kim and Nuraly K. Avliyakulov contributed equally to this study. We thank the Dean’s Office of the David Geffen School of Medicine at UCLA and Senior Associate Dean Leonard Rome for the generous and continuing support of this work.
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