In HeLa cells the expression of the BAG-3 gene, a member of the BAG family, is regulated by heavy metals and temperature, with kinetics of accumulation of mRNAs similar to Hsp70 and metallothioneins. Western blot assays performed with a polyclonal anti-BAG-3 antibody confirmed that higher levels of the protein were present in the cells following heat and metal exposure. By immunofluorescence techniques and cell fractionation assays we demonstrated that following stress BAG-3 protein concentrated in the rough endoplasmic reticulum and associated with the heavy membrane fraction. The role of BAG-3 protein during apoptosis and cellular stress is discussed.