Rhodamine 110 proved to be a very suitable label for fluorescent protease assays. The red-shifted excitation (492nm) and emission (529nm) shows better signal-to-noise ratio and produces less interference with potential inhibitors in screening assays. This leads up to a 300-fold higher sensitivity when compared with AMC substrates.
Rhodamine 110 Label
Symmetric & Asymmetric Rhodamine 110 Substrates
Due to the two amino groups of the rhodamine 110, two different types of substrates can be generated: symmetric or asymmetric rhodamine substrates. In contrast symmetric substrates, one site of the rhodamine 110 is blocked by a non-cleavable D-proline or γ-glutamate.
Compared to symmetric Rh110-substrates, asymmetric substrates offer significantly simpler kinetics upon analysis. Due to the blocking group, the entire reaction terminates after one cleavage step, as opposed to symmetric Rh110-substrates which undergo a second proteolytic cleavage step involving more complex kinetics.
Target & Substrates
BIOSYNTAN offers custom tailored rhodamine 110 substrates of symmetric and asymmetric type. A small selection of substrates is listed in the table below. For the design and synthesis of different substrates, please inquire.
| Target | Substrate |
|---|---|
| caspase-3 | Z-DEVD-Rh110-(D-Pro) |
| MALT1 | Ac-LRSR-Rh110-(D-Pro) [1] |
| calpain | Succ-LLVY-Rh110-(D-Pro) |
| trypsin, prostatin, matriptase | Bz-QAR-Rh110-(D-Pro) |
Literature
[1] C.Wiesmann et al.; J. Mol. Biol. 2012, 419, 4-21.