| Assay Method Information | |
| | Evaluation of Compound Inhibitory Activity (IC50) |
| Description: | Briefly, compounds are serially diluted in DMSO (working dilutions). ASF-coated 384 well plates are supplemented with 22.8 uL/well of biotinylated hGal-3 or hGal-1 in assay buffer (i.e. 300-1000 ng/mL biotinylated hGal-3 or hGal-1) to which 1.2 uL of compound working dilutions are added and mixed. Plates are incubated for 3 hours at 4 C., then washed with cold assay buffer (3x50 uL), incubated for 1 hour with 25 uL/well of a streptavidin-peroxidase solution (diluted in assay buffer to 80 ng/mL) at 4 C., followed by further washing steps with assay buffer (3x50 uL). Finally, 25 L/well of ABTS substrate is added. OD (410 nm) is recorded after 30 to 45 min and IC50 values are calculated. |
| Affinity data for this assay | |
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