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Gut metabolite analysis After accurately measuring the 50 mg amount of rat feces sample ( n = 6), it was transferred to a 2 mL EP tube, followed by the addition of 600 L of a methanol solution containing 4 ppm of 2-chloro-L-phenylalanine, which had been stored at -20 C

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Biotinylated goat anti-mouse IgG or goat anti-rabbit IgG (Vector Labs, BA-9200 1:200 and BA-1000 1:200, respectively) secondary antibodies were added in blocking buffer and incubated for 1 h at room temperature

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