I am investigating the properties of ADAM-17 substrate (HY-P10174) and would like to know if they exhibit any background fluorescence. Additionally, what is the underlying principle behind their detection? Any insights or references on this topic would be greatly appreciated!
It's a fluorogenic substrate that is self-quenched. The fluorophore is on the N-terminus of the peptide, and the quencher is on the C-terminus. Internal cleavage of the peptide by the protease separates the fluorophore from the quencher, resulting in an increase in fluorescence. There is usually some background fluorescence of uncleaved peptides of this sort, but a several-fold increase in fluorescence upon cleavage. This system allows for continuous monitoring of peptide cleavage using a spectrofluorimeter or fluorescence plate reader.
Article New Fluorogenic Substrates forN-Arginine Dibasic Convertase
ADAM-17 Substrate is a fluorescence-quenching peptide substrate based on the fluorescence resonance energy transfer (FRET) mechanism. Abz is a fluorophore and Dnp is a quencher. Background fluorescence is relatively weak, but enzymatic cleavage releases the quencher, increasing fluorescence intensity (Ex/Em = 330/420 nm).
The answer to this question comes from MedChemExpress Technical Support.
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