RA production assays where the amount of RA produced from a given substrate over a defined time typically utilizes 1 × 10 5 cells (or greater, if available) per replicate. Alternatively, assessment of RA production is also a useful metric for evaluating RA homeostasis. However, amounts of RA can be low in cellular systems necessitating large numbers of cells for analysis (e.g., 1 × 10 6 cells or greater depending on the cell type). If basal RA level determination is desired, skip to part 3 in Section 2.2 under “Protocols,” Retinoid Extraction. Trainor, in Methods in Enzymology, 2020 2 Direct quantification of retinoic acid production and endogenous retinoid levels in cellsīasal levels of RA can be determined in primary or cultured cells.
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