A luminescent-based protocol for NAD + /NADH detection in C. elegans, mice, and human whole blood.

Here, we present a NAD ⁺ /NADH detection assay for evaluating NAD ⁺ , NADH, and NAD ⁺ /NADH ratio across diverse biological models, including Caenorhabditis elegans, mouse muscle tissue, mouse whole blood, and human whole blood. We describe steps for sample collection and preparation from different models as well as detection and calculation of NAD ⁺ and NADH levels. This protocol is applicable for quantifying cellular/tissue NAD ⁺ and NADH levels across different biological models.
Competing Interests: Declaration of interests E.F.F. is a co-owner of Fang-S Consultation AS (organization number 931 410 717) and NO-Age AS (organization number 933 219 127); he has an MTA with Lmito Therapeutics Inc. (South Korea), a CRADA arrangement with ChromaDex (USA), and a commercialization agreement with Molecule AG/VitaDAO; and he is a consultant to MindRank AI (China), NYO3 (Norway), and AgeLab (Vitality Nordic AS, Norway). In the past three years, C.A.L. has consulted for Astellas Pharmaceuticals, Odyssey Therapeutics, Third Rock Ventures, and T-knife Therapeutics and is an inventor on patents pertaining to Kras-regulated metabolic pathways, redox control pathways in pancreatic cancer, and targeting of the GOT1-ME1 pathway as a therapeutic approach (US Patent No: 2015126580-A1, 05/07/2015; US Patent No: 20190136238, 05/09/2019; International Patent No: WO2013177426-A2, 04/23/2015).
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