# | Aladdin ID | Assay Type | Description | Organism | Compounds | Reference | BAO Format | Source | |
---|---|---|---|---|---|---|---|---|---|
1. | ALA3802986 | Binding | Inhibition of BALB/c mouse 5'-His6-tagged 3'-FLAG-tagged FAP fused with ECD expressed in HEK293 cells using Ala-Pro-p-nitroanilide as substrate preincubated for 15 mins followed by substrate addition measured for 10 mins by spectrofluorimetry | Mus musculus | 5 | ALA3797179 | cell-based format | Scientific Literature | |
2. | ALA997753 | Binding | Activity of mouse FAP expressed in HEK293 cells assessed as enzyme-mediated drug cleavage after 24 hrs by confocal microscopy relative to control | Mus musculus | 1 | ALA1140867 | cell-based format | Scientific Literature | |
3. | ALA997755 | Binding | Activity of mouse FAP expressed in HEK293 cells assessed as enzyme-mediated drug activation by measuring intracellular fluorescence after 24 hrs by confocal microscopy relative to control | Mus musculus | 1 | ALA1140867 | cell-based format | Scientific Literature | |
4. | ALA997756 | Binding | Activity of mouse FAP expressed in HEK293 cells assessed as enzyme-mediated drug activation by measuring intracellular fluorescence after 1 hr by confocal microscopy relative to control | Mus musculus | 1 | ALA1140867 | cell-based format | Scientific Literature | |
5. | ALA997757 | Binding | Activity of mouse FAP expressed in HEK293 cells assessed as enzyme-mediated drug activation by measuring intracellular fluorescence after 6 hrs by confocal microscopy | Mus musculus | 2 | ALA1140867 | cell-based format | Scientific Literature | |
6. | ALA997759 | Binding | Ratio of Kcat to Km for mouse FAP assessed as enzyme mediated drug cleavage by measuring increase in fluorescence intensity | Mus musculus | 1 | ALA1140867 | single protein format | Scientific Literature | |
7. | ALA997764 | Binding | Binding affinity to mouse FAP assessed as enzyme mediated drug cleavage by measuring increase in Pyro fluorescence | Mus musculus | 1 | ALA1140867 | single protein format | Scientific Literature | |
8. | ALA1002880 | Binding | Activity of mouse FAP assessed as enzyme mediated drug cleavage by measuring increase in fluorescence intensity | Mus musculus | 1 | ALA1140867 | single protein format | Scientific Literature | |
9. | ALA1003612 | Binding | Binding affinity to mouse FAP assessed as enzyme mediated drug cleavage by measuring Pyro fluorescence | Mus musculus | 2 | ALA1140867 | single protein format | Scientific Literature | |
10. | ALA2026780 | Binding | Inhibition of mouse recombinant FAP expressed in HEK293 cells assessed as pNA release from Ala-Pro-p-nitroanilide pre-incubated with enzyme for 15 mins prior to substrate addition by fluorescence technique | Mus musculus | 40 | ALA2021822 | cell-based format | Scientific Literature | |
11. | ALA2185388 | Binding | Inhibition of mouse recombinant FAP expressed in HEK293 cells assessed as pNA release from Ala-Pro-p-nitroanilide pre-incubated with enzyme for 15 mins prior to substrate addition by fluorescence technique | Mus musculus | 32 | ALA2176903 | cell-based format | Scientific Literature | |
12. | ALA2345663 | Binding | Inhibition of mouse FAP expressed in human HEK293 cells using N-terminal blocked Z-GP-AMC substrate at 0.036 uM to 1 uM preincubated for 10 mins prior to substrate addition by fluorescence assay in presence of prolyl endopeptidase | Mus musculus | 1 | ALA2331146 | cell-based format | Scientific Literature | |
13. | ALA2389863 | Binding | Inhibition of mouse recombinant FAP expressed in HEK293 cells using Ala-Pro-p-nitroanilide as substrate incubated for 15 mins prior to substrate addition | Mus musculus | 35 | ALA2385017 | cell-based format | Scientific Literature | |
14. | ALA3239280 | Binding | Inhibition of recombinant mouse FAP purified from HEK293 cell supernatant using Ala-Pro-p-nitroanilide as substrate by spectrophotometry | Mus musculus | 60 | ALA3232967 | single protein format | Scientific Literature | |
15. | ALA3239286 | Binding | Irreversible inhibition of recombinant mouse FAP purified from HEK293 cell supernatant using Ala-Pro-p-nitroanilide as substrate by kinetic study | Mus musculus | 1 | ALA3232967 | single protein format | Scientific Literature | |
16. | ALA3239069 | Binding | Reversible inhibition of recombinant mouse FAP purified from HEK293 cell supernatant using Ala-Pro-p-nitroanilide as substrate by kinetic study | Mus musculus | 3 | ALA3232967 | single protein format | Scientific Literature | |
17. | ALA3888519 | Binding | Enzymatic Assay: Enzyme activities were determined kinetically in a final volume of 200 μl for 10 minutes at 37° C. by measuring the initial velocities of pNA release (405 nm) from the substrate using a Spectramax plus microtiterplate reader (Molecular devices). One unit of enzyme activity was defined as the amount of enzyme that catalyzes the release of 1 μmol pNA from the substrate per minute under assay conditions. The chromogenic substrate Gly-Pro-p-nitroanilide (100 μmol/l) was used at pH 8.3 for DPP IV, Lys-Ala-p-nitroanilide (1 mmol/l) at pH 5.5 for DPP II, Ala-Pro-p-nitroanilide (300 μmol/l) at pH 7.4 for DPP9 and Ala-Pro-p-nitroanilide (2 mmol/l) at pH 7.4 for FAP activity measurement. To evaluate the endopeptidase activity of FAP and the influence of inhibitors thereon, Z-Gly-Pro-AMC and Z-Gly-Pro-p-nitroanilide were used at a final concentration of 300 and 100 μmol/l, respectively. The substrate concentrations were chosen around the Km value obtained under the assay conditions used. Buffer compositions for the DPP assays were reported before in the purification articles−vide supra. The FAP assay buffer consisted of 50 mM Tris pH7.4 containing 100 mmol/l NaCl and 0.1 mg/ml bovine serum albumin. The PREP activity was measured as described by Brandt et al. using the chromogenic substrate Z-Gly-Pro-p-nitroanilide (0.25 mmol/l) at pH 7.5 in the presence of 10 mmol/l DTT. Test compounds were dissolved and diluted in DMSO (final concentration DMSO during assay 5% v/v) except for FAP where dilution of the inhibitor was done in water. Inhibitors are pre-incubated with the enzyme for 15 min at 37° C. before starting the assay by the addition of substrate. The concentration of enzyme and of inhibitor during the preincubation is double of the final concentration during activity measurement. | 49 | ALA3886746 | single protein format | BindingDB Database |