TLC demonstration that NADH and GSH reduce NBT to the same formazan product with different kinetics.

Arrowheads show the origin and final solvent front; UV light was absorbed in the TLC plate by NADH and NBT at the origin, and by diformazan at the arrow. Reduced formazan had an Rf value ranging from 0.42 to 0.44 [38] depending on whether the lane was in the middle or edge of the TLC plate. In the absence of enzyme, GSH reduced NBT after 60 minutes but NADH only reduced NBT after 24 hours. Diaphorase or GR was necessary and sufficient for NADH to reduce NBT within 1 hour. See (S1 File) for original TLC plate images. (TIF)