| Assay Method Information | |
| | ATP-Glo Assay |
| Description: | 1. Diluted compounds in DMSO by hand for 11 points, 3 folds dilution. Then transferred 0.02 μL compounds to 384 assay plate by ECHO.2. Added 2 μL specified concentration of ENPP1 to 384 assay plate. Centrifuged 1000 RPM for 1 min.3. Added 2 μL specified concentration of ATP to the assay plate. Centrifuged 1000 RPM for 1 min.4. Incubated at 25° C. for 60 min.5. Added 4 μL AMP-Glo Reagent to the assay plates.6. Centrifuged 1000 RPM for 1 min, incubate at 25° C. for 1 hours.7. Added 8 μL Kinase Detection Reagent to the assay plates.8. Centrifuged 1000 RPM for 1 min, incubated at 25° C. for 1 hours. The final assay reaction mixture contained a buffer of 50 mM Tris pH 8.8, 250 mM NaCl and 0.1% BSA9. Read on Envision for US LUM as RLU.10. Analyzed the raw data using the equation (V. Data analysis). |
| Affinity data for this assay | |
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