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A: The structures of the Hh active oxysterols 24S-OHC, 25-OHC, and 27-OHC. B: Charge tagging of 24S-OHC with Girard T reagent. In step 1, the 3β-hydroxy-5-ene is enzymatically oxidized to 3-oxo-4-ene with cholesterol oxidase (1 h, 37°C). In step 2, the Girard T reagent reacts with the 3-oxo group to form Girard T derivates of oxysterol. See Materials and Methods for more details.
