| Assay Method Information | |
| | Gel-based Fluorescence Assay |
| Description: | The oximes were evaluated by gel-based TDP1 fluorescence assay in a concentration of 100 μM in DMSO. The fluorescence of DMSO blank vial was set as 0 and the fluorescence for the reference without TDP1 was set as 100%. Preparation: A mixture of aminooxy-containing 105 or 106 (10 μL, 30 mM in DMSO), aldehydes M1-T12 (10 μL, 30 mM in DMSO) and acetic acid (10 μL, 150 mM in DMSO) were agitated at room temperature overnight. Oximes 5-M1-T12 or 6-M1-T12 (30 uL, 10 mM in DMSO) were afforded. Two sets of oximes 105-X and 106-Y (XZ700-M1-T12, XZ699-M1-T12) (20 uL, 10 mM in DMSO) were prepared about 240 each starting from aminooxy-containing 105 (XZ700) or 106 (XZ699) based on the following method. A mixture of aminooxy-containing 105 or 106 (10 μL, 30 mM in DMSO) with about 240 aldehydes M1-T12 (10 μL, 30 mM in DMSO) (Table 4) separately in the present of acetic acid (10 μL, 150 mM in DMSO) were agitated at room temperature overnight in three 96-well plates in parallel. The formed oximes 105-M1 to 105-T12 and 106-M1 to 106-T12 (30 uL, 10 mM in DMSO) were diluted to 100 μM in DMSO and evaluated by gel-based TDP1 fluorescence assay. The fluorescence of the gel band of DMSO blank vial was set as 0 and the fluorescence for the reference gel band without TDP1 was set as 100%. |
| Affinity data for this assay | |
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