Nad nadh colorimetric kit

Cells transfected with either the NBN full-length- or the p26-coding vector were treated with 2 Gy of X-rays and harvested after 0.5, 2, and 24 h. Briefly, 2×10⁵ cells were washed in PBS and lysed. To remove NADH-consuming enzymes, the extracted samples were filtered through 10 kDa molecular weight cut off filters (Abcam) before performing the assay. The NAD⁺/NADH quantification was performed using the NAD⁺/NADH colorimetric kit, according to manufacturer's instruction (BioVision, Milpitas, USA). Samples were read at OD 450 nm, using the microplate reader Victor X (PerkinElmer, Waltham, USA).

The amounts of ATP and the ATP/ADP ratios were measured with an ADP/ATP ratio assay kit (Abcam) according to the manufacturer's instructions. The amounts of ATP were determined by an ATP standard curve, and normalized to the protein concentrations. The amounts of NADH and NADH/NAD ratios were measured with an NAD/NADH colorimetric kit (BioVision). The NADH concentrations were determined by a NADH standard curve, and normalized to the protein concentrations.