Notes: Human MTHFD2, a mitochondrial methylenetetrahydrofolate dehydrogenase and cyclohydrolase, and the dehydrogenase (D) and cyclohydrolase (C) domain of MTHFD1 were purified from bacteria. To determine the IC₅₀ values of LY345899, MTHFD2 and MTHFD2 reactions proceeded for 15 minutes before NAD(P)H-Glo™ Detection Reagent was added to all wells in the plate, incubated for 60 minutes and luminescence measured. (4961)

Notes: The reducing activity of purified recombinant human Apurinic apyrimidinic endonuclease 1/Redox factor-1 (rhAPE1/Ref-1) was quantitated using the NAD(P)H-Glo™ Detection System. (4962)

Notes: NIH3T3 cells were grown in medium containing 2% or 8% FBS for 24 hours and then NADH and NADPH levels assessed using the NAD(P)H-Glo™ Detection System. (4960)

Notes: Cell pellets of A375 human melanoma cells were lysed by passing through a needle, centrifuged to remove debris and protein quantitated. Twenty micrograms of protein from control and treated lysates were digested with 1µg of Sequencing Grade Modified Trypsin and used for mass spectrometry analysis. A375 cells (5 × 10⁵) were grown in a 10cm dish and treated for 24 hours before isolating RNA and DNA synthesized using M-MLV Reverse Transcriptase. The cDNA was then used in qPCR. A375 cells were plated at 3 × 10³ in 96-well half-volume white-wall plates, grown and treated for 24 hours. NAD, NADH and NADPH were determined using the NAD(P)H-Glo™ Detection System or NAD/NADH-Glo™ Assay and luminescence measured. (4963)