Nadp nadph quantitation kit

For NADPH measurements, C. crenatum cells were harvested at the exponential phase during the batch fermentations in shake flasks by centrifugation (5,000 g for 10 min at 4 °C). The intracellular concentrations of NADPH were measured using a NADP/NADPH quantitation kit (BioVision, Inc., Milpitas, CA) according to the manufacturer’s instructions.
Total RNA was extracted from C. crenatum cells at the exponential phase during the batch fermentations in shake flasks using the RNAiso Plus reagent (Takara, Dalian, China). The cDNA was synthesized with a PrimeScript RT reagent kit (Takara, Dalian, China). Real-time PCR (RT-PCR) was performed on a Bio-Rad CFX96 Touch Real-Time PCR Detection System (Bio-Rad Hercules, CA) using the SYBR Premix Ex Taq^TM II (Takara, Dalian, China). The 16S rRNA gene was used as the internal standard16 (link). The primer sequences used in RT-PCR are supplied in the supplementary section (Table S18). Each assay was replicated three times.

Fly heads of each genotype were collected by being frozen in liquid nitrogen. Thirty frozen fly heads were washed with cold PBS and homogenized with 150 µl NADP/NADPH extraction buffer provided by NADP/NADPH quantification kit (BioVision, Cat.# K347‐100). NADPH concentration of each sample was measured according to the protocol in NADP/NADPH quantitation kit (BioVision, Cat. # K347‐100) and was normalized by its own total protein (Pierce BCA protein assay kit, Prod # 23227). At least three independent samples were analyzed for NADPH measurement.

The colorimetric NADP/NADPH Quantitation Kit (Biovision Inc., Milpitas, CA, USA) was used to detect the NADPH levels. The absorbance at 450 nm was read and normalized to the number of cells.