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Perform small pilot titrations to identify the linear response window for your assay rather than assuming a single working concentration

This is when the effects are strongest

Dihydroorotate dehydrogenase is involved in pyrimidine synthesis and uses riboflavin as flavin mononucleotide (FMN) instead of FAD, while mitochondrial glycerol 3-phosphate dehydrogenase depends on FAD like all the red enzymes