NADP⁺ and NADPH measure
Description
The abundance of NADP+ and NADPH as well as the ratio of NADP+/NADPH measured by the LC-MS analysis in naïve NANOG iKO SHhES8 hESCs, either untreated or treated with DOX for three days. Data are presented as mean ± SEM (n=6).
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Steps to reproduce
For NADP+ and NADPH quantification, naïve hESCs, either without or with DOX treatment for 3 days, were mechanically collected with 1 mL of methanol: acetonitrile: water (2:2:1, v/v) solution. Samples were analyzed by the Applied Protein Technology (Shanghai, China). The method is the same as previously reported. Briefly, following the SRM/MRM method, metabolite analysis of NADP+ and NADPH was performed on a system consisting of an UHPLC (1290 Infinity LC, Agilent) coupled to a QTRAP mass spectrometer (AB SCIEX 5500). Quantitative information from every sample set was extracted in the Multiquant Software.
Institutions
- Shanghai Jiao Tong University School of Medicine