Heat shock protein 70 (Hsp70) family members are known as facilitators of immune responses by interacting with receptors on antigen-presenting cells leading to Hsp70-peptide uptake and antigen cross priming. Here, identification of human leukocyte antigen (HLA)-A24-restricted epitopes was achieved using peptide arrays for evaluation of their affinities to Hsp70 and HLA-A24 binding prediction tools. Using Hsp70 as the model antigen, the GYPVTNAVI and VFQHGKVEI peptides were identified as antigens. These peptides actually bound to HLA-A24 in the stabilization assay using T2-A*2402 cells, and induced a strong peptide-reactive cytotoxic T lymphocyte response in HLA-A24 transgenic mice after vaccination.