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.
Department of Chemistry and Biochemistry, University of California Los Angeles, Los Angeles, CA, USA.
Michael E
ME
Jung
Wan-Joong
WJ
Kim
Nuraly K
NK
Avliyakulov
Merve
M
Oztug
Michael J
MJ
Haykinson
D002232:Carbocyanines / Q000138:chemical synthesis* / Q000737:chemistry; D060326:Chemistry Techniques, Synthetic / Q000379:methods*; D004396:Coloring Agents / Q000138:chemical synthesis* / Q000737:chemistry; D004355:Drug Stability; D004356:Drug Storage; D007525:Isoelectric Focusing; D015203:Reproducibility of Results; D013997:Time Factors; D057825:Two-Dimensional Difference Gel Electrophoresis / Q000379:methods*
D002232:Carbocyanines; D004396:Coloring Agents