| Assay Method Information | |
| | ENPP1 Enzyme Inhibition Assay |
| Description: | Table 2, Column 1: ENPP1 catalyses the hydrolysis of ATP into AMP+PPi. AMP production was monitored using Bellbrooks Transcreener AMP/GMP TR-FRET assay kit (Cat No. 3020-10K) according to the manufacturer's instructions. The assay was run with 50 mM Tris pH 7.5, 5 mM MgCl2, 150 mM NaCl, 0.005% Tween20 and distilled H2O. Antibody and tracer were diluted to 2 nM. Human ENPP1 was run at 20 pM with ATP at 300 nM. Mouse ENPP1 was run at 15 pM with ATP at 400 nM. AMP produced in the assay by ENPP1 activity competes with a far-red tracer labelled AMP bound to an AMP specific antibody resulting in a decrease in TR-FRET. ENPP1 inhibition reduces AMP production and increases the TR-FRET signal. |
| Affinity data for this assay | |
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