| Assay Method Information | |
| | Human LOXL2 Purified Recombinant Protein Assay |
| Description: | Table 9: The amine oxidase activity was evaluated by measuring Amplex Red fluorescence using commercially available purified, recombinant human LOXL2 (Sino Biologicals, Beijing, China). To assay for amine oxidase activity, 10 μL of 0.025 μg/μL purified, recombinant LOXL2 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) are added and the plate mixed and immediately placed on the FlexStation for fluorescence measurements. Fluorescence is read in kinetic mode every 2 min for 0.5-1 hour at excitation=544 and emission=590. The amine oxidase activity is 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 βAPN (3-aminopropionitrile) represented no activity and were set to 100% inhibition. |
| Affinity data for this assay | |
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