Microfluidic paper-based analytical devices (μPADs) have attracted much attention because of their various advantages such as small size, ease of fabrication, low cost, and minimum requirement of sample volume for measurement. However, not much has been given to the development of flow control techniques. Therefore, in this study, we developed a temperature responsive valve that can be applied to μPADs as a flow control method of μPADs by applying poly(N-isopropylacrylamide) to PVDF membrane, and demonstrated valve function to protein which is closed and opened by controlling temperature at 25 °C and 40 °C, respectively.