Educational guide
How to prepare the Cy3 and Cy5 labeled peptides? | LifeTein Peptide Blog
Peptides were synthesized using standard Fmoc solid-phase peptide synthesis. After coupling, Lys(Mtt) was selectively deprotected to expose the side-chain amine moiety. Cy3 or Cy5 were dissolved in DMF, activated, and coupled to the free amine. Cy3—and Cy5-lab
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Peptides were synthesized using standard Fmoc solid-phase peptide synthesis.
After coupling, Lys(Mtt) was selectively deprotected to expose the side-chain amine moiety. Cy3 or Cy5 were dissolved in DMF, activated, and coupled to the free amine. Cy3—and Cy5-labelled peptides were cleaved and HPLC purified according to the standard procedure.
Lyophilization yielded a red powder for Cy3 and blue for Cy5.
Stock solutions of non-labeled peptide (10 mM), Cy3-labelled peptide (1 mM), and Cy5-labelled peptide (1 mM) were prepared in water, aliquoted, and stored at −20 °C.
Excitation wavelengths of 520 or 615 nm for Cy3 or Cy5, respectively, were used.
Use emission wavelengths of 600 or 710 nm for Cy3 or Cy5, respectively.