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The membranes were blocked with 5% nonfat milk in Tris-buffered saline containing 0.1% Tween-20 for 1 h at 37 C and then incubated with primary antibodies overnight at 4 C

Overgrowth of bacteria: Some people develop vitamin B 12 deficiency as a result of conditions that slow the movement of food through the intestines (diabetes, scleroderma, strictures, diverticula), allowing intestinal bacteria to multiply and overgrow in the upper part of the small intestine