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Free glutathione, GSH, is read first after 15 minutes, followed by addition of a reaction mixture that converts all the oxidized glutathione, GSSG, into free GSH, which then reacts with the excess ThioStar to yield the signal related to Total GSH content

It's often helpful to draw a small amount of air into the syringe first, inject that air into the vial (to equalize pressure), and then draw the liquid