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The pyloric valve, which separates the stomach from the small intestine, closes when food is present in the stomach (especially protein and fatty foods)

The PVDF membrane was subsequently washed (5 min 5) with tris-buffered saline with tween (TBST) buffer and then incubated with the appropriate secondary antibody (Peroxidase-conjugated goat anti-rabbit IgG (H + L), at a dilution of 1:5000) in a 5% bovine serum albumin solution for 1 h at 25 C