The new store with be the family-run company’s eleventh store since it began in Stockton on Tees in 1946.
The £1m investment will see an existing building in Old Shoreham Road, Hove, be converted to offer 35,000 sq ft of space across two floors.
The firm has already boosted its Southern portfolio with a showcase store in Battersea, London, which opened in 2013, and work is currently underway on a store in Guildford, Surrey.
James Barker, managing director of Barker and Stonehouse, said: “Our plans to expand into the South East are really taking shape and we’re delighted to be able to announce this latest venture.
“The Brighton and Hove store will be designed to be somewhere people like to visit, with a breadth of products and the quirky touches that make shopping in Barker and Stonehouse an exciting customer experience.
“Barker and Stonehouse has built up a loyal following at each of its stores due to the wide choice of quality products and the excellent customer service offered by its very knowledgeable staff.”
The store is set to open in spring 2017.