I am trying to measure intracellular (in vitro) NAD using EnzyChrom NAD assay kit, however the results are variable. I was wondering if cell dissociation could affect its level. Are there tips and tricks needed or is there another more accurate method to measure NAD level?
S. Béatrice Marianne Ewalds-Kvist
Sample Preparation: Cells or tissues are lysed to release NAD. Extraction: NAD is extracted from the lysate using a suitable solvent. Separation: The extract is then separated using HPLC. Detection: The separated NAD is detected and quantified using LC-MS/MS.
The most accurate method to measure intracellular NAD levels is High-Performance Liquid Chromatography (HPLC) coupled with tandem mass spectrometry (LC-MS/MS). This method provides highly sensitive, reliable, and reproducible results1. Here's a brief overview of the process:
This method allows for precise quantification of NAD levels and is widely used in research settings.
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