Nicotinamide adenine dinucleotide (NAD) is vital to many cellular processes and is distributed between distinct subcellular pools in the compartmentalized eukaryotic cell. The detection and relative quantification of these individual pools is difficult because of the methods usually applied, which require cell disruption and fractionation.
Here, we describe an immunochemical method to visualize and relatively quantify subcellular NAD+ pools, which relies on the NAD+-consuming activity of poly-ADP-ribose polymerase 1 (PARP1). We demonstrate that this system can be readily applied to detect changes in the mitochondrial, Golgi, endoplasmic reticulum, and peroxisomal NAD+ pools.
Compartmentalization Mitochondria PARP1 Poly-ADP-ribose NAD Biosensor Immunocytochemistry Immunoblot analysis
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MVL, MN, MZ, and CD were supported by the Research Council of Norway (project #. 214435). A.A.N. was supported by the Russian Science Foundation (Grant # 16-14-10240).
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