Incubation of the cytosolic S100 cell extract with dATP and Cyt c restored the apoptotic pathway through induction of the apoptosome formation (Fearnhead, 2001); this restoration was followed using a fluorogenic substrate for caspases (Ac-DEVD-afc).
To examine the content of apical proteins in the channel membranes, the bile canalicular enzyme dipeptidyl peptidase IV (DPPIV) was localized using a fluorogenic substrate, Ala-Pro-cresyl violet.
By using a fluorogenic substrate, Z-Arg-Leu-Arg-Gly-Gly-AMC, the deubiquitinating activities for H6YUH1 (1.72 U/mg) and YUH1H6 (1.61 U/mg) were about 18 times higher than 0.092 U/mg for H6UBP1, ubiquitin specific protease 1 of S. cerevisiae containing the 6 histidine residue at the N-terminus which is normally used in protein engineering.
We used a fluorogenic substrate for the β-galactosidase enzyme to quantify enzymatic activity in a Wallac Victor 3 fluorimeter (Methods).
Using a fluorogenic substrate, we did detect a slight decrease in 20S proteasome activity in vitro, but not in 26S proteasome activity – although 20S proteasome does exist in cells, it is thought that the 26S proteasome is responsible for the degradation of ubiquitylated proteins [37].
The activity was measured using a fluorogenic substrate (Fluorescent FXIII Assay; Zedira GmbH, Darmstadt, Germany).
Using a fluorogenic substrate, we demonstrated that GST Sir2 is a catalytically active lysine deacetylase.
Media samples were analysed for total MMP activity using a fluorogenic substrate assay.
α-Secretase activity was measured using a fluorogenic substrate (10 μg/reaction, alpha-secretase substrate II; Merck, Whitehouse Station, NJ, USA), as indicated by the manufacturer.
The in situ zymography allowed identification of the gelatinolytic activity, due to expression of MMPs on frozen sections of unfixed masseter muscle samples using a fluorogenic substrate (gelatine).
