I am trying to establish the in vitro ubiquitination to screen the compounds,the reaction system(E1,E2,ATP and reaction buffer) is the same as generally used,and the unreacted(remaining) ATP was determined wih CellTiter-Glo® Luminescent Cell Viability Assay(Promega)(my mentor adviced me to do so) following the manufacture' instructions,however,I can not get consistent results(in fact,the results may vary significantly from time to time),I wonder if it is the intrinsic disadvantage of the methods or there is some tricks.Does anyone use similar methods,or give me some advice?Any suggestion is welcome.

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