Organ culture and IgA ELISA

Bladders were aseptically removed, bisected, rinsed with PBS, and both halves cultured together in 500 μL RPMI-1640 with 10% FBS, 1% Pen/Strep (15–140-122, Gibco), 10 mM HEPES, and 1% Glutamax (35030–061, Gibco). Supernatants were removed after 24 hrs and cleared of debris by centrifugation. IgA concentration in urines and culture media was determined by ELISA according to manufacturer protocol (88–50450-22, Invitrogen).