Charles Clinkard, which was founded in 1924, has closed its site on Banbury Road in Summertown, Oxford. The reason for the closure is unclear.
Customers in Oxfordshire can still visit the Witney’s Woolgate Shopping Centre and the high street in Burford sites.
The closure of the Summertown branch comes after the brand’s Teesside Park site closure in May.
The more than 100-year-old company also closed its original Middlesbrough store in 2024.
The family-run business has previously said changing shopping habits and high street challenges had forced it to focus on improving performance across its remaining stores.
Managing director Tim Payne told Retail Week earlier this year that the company was considering closing around two-fifths of its store locations.
The Clinkard Group currently operates 32 stores across the UK in addition to its online operation.
The retailer has shifted its focus to online sales as an increased number of customers prefer online shopping.
It is not clear whether more closures are planned.