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Wheat protein-induced hypersensitivities (e.g., Celiac disease, wheat allergy, non-Celiac gluten sensitivity, etc.) are growing at an alarming rate on a global scale for reasons that are largely unknown. To advance fundamental mechanisms underlying these disorders, we developed a mouse model of hypersensitivity to alcohol-soluble proteins (ASP) isolated from Ambassador soft wheat. Wheat flour was prepared and ASP were extracted using established methods. The ASP fraction was characterized by SDS-PAGE analysis. Groups of BALB/c mice were injected with the ASP along with alum five times, at ten-day intervals. Blood collected before and after injection was analyzed for specific IgE, IgG1, IgG2a and IgA antibody (Ab) responses using optimized ELISA methods. Mice received an i.p. challenge with the ASP and were evaluated for hypothermia shock response (HSR) by rectal thermometry. Blood collected after the challenge was analyzed for murine mucosal mast cell protease (mMCP)-1 elevation. The ASP fraction contained nine major proteins (8 with sizes from 12 to 38 kDa, 1 of >170 kDa, smear) and two minor proteins (70, 128 kDa). Upon injection, mice developed time-dependent robust specific IgE, IgG1 and IgG2a, but very little IgA Ab, responses. Systemic challenge resulted in significant HSR, but mice recovered by one hour. There was a significant elevation of mMCP-1 in the blood after challenge, confirming mast cell/IgE Ab mediated anaphylactic reaction. These data demonstrate that this mouse model may be used for basic and applied studies on soft wheat gluten-induced hypersensitivity disorders.