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As global temperatures rise due to climate change, the occurrence and intensity of heat waves are anticipated to increase. Despite this anticipation, there remains a gap in comprehending the underlying defense mechanisms against heat stress, particularly exhibited by Heat Shock Proteins (HSP) within vulnerable populations. Hence, this study aims to assess the expression of heat shock protein 70 (HSP70) in response to heat exposure among vulnerable populations residing in both urban and rural areas. A total of 108 respondents were recruited using multistage sampling. This study consists of three (3) methods: Part I: face-to-face questionnaire; Part II: indoor heat exposure monitoring using Universal Thermal Climate Index (UTCI) classification; and Part III: blood sample collection to determine HSP70 protein expression using HSP70 High Sensitivity Enzyme-linked Immunosorbent Assay (ELISA) kit. The results show that urban areas were exposed to strong UTCI thermal stress, whereas rural areas experienced moderate UTCI thermal stress. A higher level of HSP70 protein expression was observed in urban (2.524 ± 0.26) than in rural respondents (2.095 ± 0.27). This study concludes that both urban and rural vulnerable populations demonstrated different patterns of HSP70 protein expression in response to various levels of heat exposure.