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After washing in PBS, primary antibodies (IgM-PE BioLegend 406507, IgD-APC BioLegend 405713, MARCO Santa Cruz sc-65353/ anti-rat PE Abcam ab97058 and Siglec-1 Biolegend 142417) diluted in PBS 10% FCS were incubated overnight at 4 C

it can survive and remain active across the highly variable pH levels of the human gastrointestinal tract, from the highly acidic environment of the stomach to the more alkaline environment of the small intestine