| Assay Method Information | |
| | Human LOXL3 Purified Recombinant Protein Assay |
| Description: | Table 9: The amine oxidase activity was evaluated by measuring Amplex Red fluorescence using commercially available purified, recombinant human LOXL3 (R&D Systems, Minneapolis, Minn.). To assay for amine oxidase activity, 10 μL of 0.075 μg/μL purified, recombinant LOXL3 diluted in Assay buffer Buffer (50 mM Borate Buffer, pH8) was incubated with 2 μL of test compound in DMSO and 73 μL Assay Buffer for 2 h at 37° C. After the 2 h incubation, 5 μL of 10 mM 1,5-Diaminopentane (DAP) diluted in Assay Buffer and 10 μL of Amplex Red Mix (8.5 μL Assay Buffer+0.5 μL of 10 mM Amplex Red+1 μL of 500 U/mL Horseradish Peroxidase) was added and the plate mixed and immediately placed on the FlexStation for fluorescence measurements. Fluorescence was read in kinetic mode every 2 min for 0.5-1 hour at excitation=544 and emission=590. The amine oxidase activity was calculated from the slope of the linear portion of the curve. Wells containing vehicle (DMSO) represented maximum activity and were set to 0% inhibition and wells containing 100 μM BAPN (3-aminopropionitrile) represented no activity and were set to 100% inhibition. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |