Even for what might be considered a straightforward assay involving fluorescent measurements of a ligand binding to a protein target, this might include (but is by no means limited to): compound impurities and degradation, imprecise compound dispensing, unmonitored water absorption by DMSO stocks, the effect of DMSO on protein stability, intrinsic compound fluorescence, compound insolubility or aggregation, variability in protein concentration or quality, pipetting errors, and inherent noise in any fluorescence measurement-not to mention stray lab coat fibers as fluorescent contaminants. This is unsurprising, given the number and variety of potential contributing factors.
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