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Assay Method Information |
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| | H6PD Activity Assay |
| Description: | HEK293 H6PD-OE and HEK293 pellets were lysed and the lysate was quantified with the Bradford protein assay. Reaction mix containing galactose-6-phosphate (Gal6P) was combined with increasing concentrations of PARPi or DMSO. As an additional control, reaction mix without substrate was combined with HEK293 H6PD-OE lysate. Lysate from HEK293 or HEK293 H6PD-OE was added to each well to start the reactions. The absorbance at 340 nM (NADPH) was measured for 90 min at 10 min intervals. The 60 min data, which were within the linear range, was used for analysis. Background absorbance was subtracted and the H6PD activity of theHEK293 H6PD-OE lysates treated with PARPi was normalized to the DMSO control. |
| Affinity data for this assay | |
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