Basic Information
ID: ALA3888127
Type: Binding
Description: In Vitro AMPK Activation Assay: The in vitro AMPK activation assay is performed in a volume of 30 μl in a 384-well plate. Enzyme reactions were assembled in the microtiter plate by adding 15 μl of 2× enzyme in assay buffer (20 mM HEPES, pH 7.3, 5 mM MgCl2, 3 mM DTT, 0.01% Brij 35 and CamK Kinase, to activate AMPK) to wells which contained either DMSO or compound. The reaction was initiated with the addition of 15 μl 2× substrate mixture containing 200 μM ATP, and 3.0 μM fluorescently labeled SAMS (5-FAM-HMRSAMSGLHLVKRR-COOH) in assay buffer. After 45-minute incubation at 25° C., the reaction was stopped by the addition of 70 μl stop buffer (100 mM HEPES, pH 7.3, 40 mM EDTA, 0.015% Brij 35). Phosphorylated 5-FAM SAMS product is assessed using a Caliper EZ Reader LabChip microfluidics reader. Product conversion is determined by calculating the peak heights of the substrate and product and reporting the product/(product+substrate) peak ratio.
Organism: Homo sapiens
Activity Charts
Bioactivity
Activity Types for Assay ALA3888127
EC50
Compound Summary
| Parent Molecular Weight | ALogP | Polar Surface Area |
|---|---|---|
| 492.89 | 3.86 | 3.86 |
| 535.98 | 4.26 | 4.26 |
| 553.02 | 2.46 | 2.46 |
| 532.94 | 2.73 | 2.73 |
| 630.97 | 3.13 | 3.13 |
| 521.02 | 4.70 | 4.70 |
| 489.96 | 5.06 | 5.06 |
| 538.00 | 3.09 | 3.09 |
| 533.93 | 3.19 | 3.19 |
| 531.96 | 3.33 | 3.33 |
| 535.00 | 3.51 | 3.51 |
| 479.92 | 3.34 | 3.34 |
| 556.02 | 4.39 | 4.39 |
| 529.98 | 4.10 | 4.10 |
| 532.98 | 3.45 | 3.45 |
| 480.95 | 2.54 | 2.54 |
| 519.99 | 4.14 | 4.14 |
| 543.97 | 4.01 | 4.01 |
| 557.99 | 3.91 | 3.91 |
| 531.96 | 4.21 | 4.21 |
| 531.96 | 2.89 | 2.89 |
| 537.02 | 3.79 | 3.79 |
| 545.00 | 3.83 | 3.83 |
| 531.96 | 3.33 | 3.33 |
| 560.99 | 3.89 | 3.89 |