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Application Note: Evaluation Of PARP Inhibitors: Performed On BMG LABTECH's FLUOstar Omega
Poly ADP-ribosylation is a post-translational modification of proteins that plays a crucial role in regulating DNA repair. Poly (ADP-ribose) polymerase (PARP) transfers ADP-ribose to itself and other nuclear proteins such as histones. The substrate for that reaction is NAD+. PARP inhibition has been demonstrated to potentiate the cytotoxicity of anti-cancer drugs and ionising radiation. Therefore much effort has been put into the development of specific PARP inhibitors. To evaluate such inhibitors, we have first used a specific PARP activity assay, to monitor their ability to inhibit endogenous PARP activity contained within a colon cancer cell line. This assay requires protein concentration determination (BCA assay - absorbance) followed by luminescent detection. Secondly, we have used the AlamarBlue viability assay (fluorescent read-out) to evaluate the ability of a PARP inhibitor to potentiate the effects of the chemotherapeutic agent temozolomide to stimulate cell death in a colon cancer cell line. All measurements were performed using a FLUOstar Omega multidetection microplate reader from BMG LABTECH.
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