| Assay Method Information | |
| | Fluorescence Direct Binding Assay |
| Description: | 1) Optimization of measurement parameters to minimize protein consumption and to minimize the dilution effect and the DMSO content2) Titration measurements of the protein against ligand by at least 12 titration steps to obtain a good s-curve fit3) Repeat the same titration measurements with the ligand alone to enable correction4) Check the stability of the protein once by titration against DMSO alone5) Determination of the molar extinction coefficients of the ligand at 280 and 340 nm with help of an UV-spectrophotometer6) Use Excel template for the correction of the measured raw data7) Use GraphPad Prism software for the quadratic binding fit and the KD evaluation. |
| Affinity data for this assay | |
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