If N-(Aminoethyl)-8-naphthylamine-1-sulfonic Acid is incorporated into a small peptide can it be used to trace the peptide by its own u/v fluorescence or is it only of use for FRET applications?
N-(Aminoethyl)-8-naphthylamine-1-sulfonic Acid (AEDANS)-labeled peptide can be checked with its optical property (UV/ fluorescence). The excitation wavelength for AEDANS-peptide can be used in 295 nm or 375 nm, and you can check the emission intensity at 484 nm. AEDANS is also of use as a donor for FRET study.
You can use the peptide itself with its UV/Vis characteristics. Note that it is reported that 1,8 ANS has increase in fluorescence only upon interaction with exposed hydrophobic patches. Is your peptide large enough to already have enough structure to have also exposed hydrophobic patches? Is your peptide flexible enoughto allow the intramolecular movement of the conjugated 1,8 ANS towards such hydrophobic patches? If not, why not use the peptide and classically add 1,8 ANS?
1,8 ANS can also be used in a FRET application, but take into account that as a donor of FRET its fluorescence quantum yield will decrease due to an acceptor in its vicinity but will also increase due to a hydrophobic microenvironment.
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