| Assay Method Information | |
| | Fluorescence Resonance Energy Transfer (FRET) Assay. |
| Description: | Assay Reaction: The assay was conducted according to the following: Each assay reaction was conducted in a volume of 25 μl in a 384-well half-volume plate. 10 point compound serial dilutions (3-fold) were generated in DMSO at a concentration of ×50 that of the final assay concentration (FAC). Compound solutions were then prepared by IgE-Tb diluting 10-fold in assay buffer. For the assay, 5 μl of diluted compound was added to 10 μl of IgE-Tb, followed by addition of 10 μl FcεR1α-Y131A-AF488. FRET reagents FACs were 5 nM IgE-Tb, 25 nM FcεR1α-Y131A-AF488. Usually the top FAC of compound in the assay was 10 μM. The final DMSO concentration was 2%. The minimum signal (MIN) was measured by adding 5 μl unlabelled FcεR1α at 1 μM (FAC=200 nM) to the FRET reagents. The maximum FRET signal (MAX) was measured in wells containing FRET reagents but no compound. The assay was incubated for 2 hours at room temperature, protected from light and evaporation, and with gentle agitation. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |