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The membrane was blocked with 5% dry skim milk in Tris-buffer saline (TBS) containing 0.05% Tween-20 (TBS-T), incubated with primary rabbit anti-GST polyclonal antibody (1:10,000) or primary mouse anti-His polyclonal antibody (1:10,000) (Proteintech Group, Chicago, USA), and then incubated with secondary alkaline phosphatase conjugated goat anti-rabbit or anti-mouse antibody (1:2,000) (Proteintech Group, Chicago, USA)

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The sample was measured at 400405 nm in multimode plate reader (Model: Enspire, Perkin Elmer, CA, USA)

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Since the absorption through the gastrointestinal tract and transfer to the thyroid gland is the primary route of iodine uptake in humans, the trillions of microorganisms in the gut thus play a critical role in regulating iodine metabolism