The pharmacy and health and beauty giant is due to open its newest branch at the retail park in Stocksbridge on March 21.
The new 6,000 sq ft store will be located beside Next and Sports Direct at the shopping complex, which opened in June 2016.
Boots' Sheffield area manager Ritesh Bakrania said: "We’re pleased to confirm that we have secured a new unit at Fox Valley, and look forward to supporting the local communities in north Sheffield with all of their health and beauty needs."
The unit had been home to the Fox Valley Friday and Saturday markets for the last three months, with the Friday market now moving back to the outdoor walkway near Home Bargains.
James Shepherd, estates director at Dransfield Properties, which owns the retail park, said: "From the feedback we have had from our shoppers we are confident the new store will be well received and will certainly bring a new offer to the centre. We look forward to the completed store opening in March."