The Cardington Road hotel has been placed on the market through hotel property specialists, Colliers International.
The freehold property is being offered to the market with offers accepted in the region of £3,500,000.
The Barns Hotel is a Grade II listed building with a 13th century Tithe Barn.
It has 49 en suite letting bedrooms and conference facilities with views of the River Ouse.
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Julian Troup, Head of UK Hotels Agency at Colliers International, said: “We are anticipating a great deal of interest from a variety of different types of buyers.
“The hotel offers the ideal range and combination of features for long term hotel operation and enjoys an enviable position close to the Dunstable Downs and also the modern city of Milton Keynes”.