1-Indanone (1-INO), 1H-phenalen-1-one (1H-PHO), acenaphthylene-1,2-quinone (1,2-ACNQ), chrysene-1,4-quinone (1,4-CHRQ), 2-methylanthracene-9,10-quinone (2-MAQ), 2,3-dimethylanthracene-9,10-quinone (2,3-DMAQ), naphthacene-5,12-quinone (5,12-NQ), 7H-benz[de]anthracene-7-one (7H-BAO), 9-fluorenone (9-FLO), phenanthrene-9,10-quinone (9,10-PQ), anthracene-9,10-quinone (9,10-AQ) and benz[a]anthracene-7,12-quinone (7,12-BAQ) all with purity ≥95% and from Sigma Aldrich (Stockholm, Sweden) were kindly provided by Dr Staffan Lundstedt at Umeå University. 4H-Cyclopenta[def]phenanthrene-4-one (4H-CPO), benzo[a]fluorenone (BFLO) and 6H-benzo[cd]pyren-6-one (6H-BPO), all with purity ≥99% were obtained from the Institute for Reference Materials and Measurements (EC-JRC-IRMM, Geel, Belgium). 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD; CASRN 1746-01-6, 98%) was obtained from Cambridge Isotope Laboratory (Andover, MA). Benzo[a]pyrene (B[a]P; CASRN 50-32-8, ≥96%), 7-ethoxyresorufin, fluorescamine, dihydronicotinamide-adenine dinucleotide phosphate (NADPH) and microsomes containing recombinant human CYP1A1 and NADPH-P450 reductase were obtained from Sigma-Aldrich (Stockholm, Sweden). All cell culture reagents were purchased from Gibco (Life Technologies, Stockholm, Sweden).
Dihydronicotinamide adenine dinucleotide phosphate nadph
Manufactured by Merck Group
Dihydronicotinamide-adenine dinucleotide phosphate (NADPH) is a coenzyme involved in various metabolic processes. It functions as a reducing agent, participating in cellular reactions that require the transfer of hydrogen atoms or electrons.
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Lab products found in correlation
Phenanthrene 9 10 quinone, by Merck Group (1 mentions)
7 ethoxyresorufin, by Merck Group (1 mentions)
1 indanone, by Merck Group (1 mentions)
Fluorescamine, by Merck Group (1 mentions)
5 sulfosalicylic acid, by Merck Group (1 mentions)
Glutathione reductase, by Merck Group (1 mentions)
Cytation 3 imaging reader, by Agilent Technologies (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Merck Group (1 mentions)
5 5 dithiobis 2 nitrobenzoic acid dtnb, by Merck Group (1 mentions)
2 protocols using dihydronicotinamide adenine dinucleotide phosphate nadph
1
Polycyclic Aromatic Hydrocarbon Compounds Protocol
2
Intracellular Glutathione Measurement Protocol
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Total intracellular glutathione measurements were performed as previously described [40 (link)]. Briefly, 5 × 105 H1299p53R273H Cas9 and H1299p53R273H-Casp2−/− cells with and without 2 µM erastin treatment for 12 h were homogenized in ice-cold 10 mM HCl. Proteins were precipitated by adding 5-sulfosalicylic acid (Sigma-Aldrich) to a final concentration of 1%. Samples were centrifuged to remove precipitates and supernatants were collected and stored at –20 °C until analysis. Samples were incubated in the presence of 5,5’-dithio-bis-[2-nitrobenzoic acid] (DTNB, 0.73 mM) (Sigma-Aldrich), EDTA (4 mM) (Sigma-Aldrich), dihydronicotinamide- adenine dinucleotide phosphate (NADPH, 0.24 mM) (Sigma-Aldrich), 110 mM sodium phosphate (NaH2PO4) buffer (pH 7.4) and glutathione reductase (GR; Sigma-Aldrich) from baker’s yeast (1.2 U/ml). The absorbance at 412 nm was measured every 15 sec for 5 min on a Cytation 3 Imaging Reader (BioTek). The total concentration of GSH + GSSG was calculated based on a GSH standard curve.
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