Microfluidic paper-based analytical devices (μPADs) have recently attracted the attention of researchers and industry owing to their various advantages. However, µPADs lack a way to control fluid flow; therefore, it is difficult to perform complex immunoassays that use multiple reagents and replace the reagents from the analytical area. We developed a controllable thermoresponsive valve for μPADs by functionalizing a polyvinylidene difluoride porous membrane with plasma-induced graft polymerization of poly(N-isopropylacrylamide) (PNIPAAm), which is a thermoresponsive polymer that changes its hydrophilic properties near the lower critical solution temperature (LCST; 32 °C). Surface analysis by attenuated total reflectance Fourier transform infrared spectroscopy confirmed that the fabricated thermoresponsive valves coated with PNIPAAm. The valve performance was evaluated by sandwiching the thermoresponsive valve between two paper microchannels stacked in a T-shaped paper microfluidic device. The thermoresponsive valve fabricated with a monomer concentration ranging from 2.3 to 3.0 wt% and a polymerization time of 5 h or 2.0 wt% and 20–22 h showed good valve performances. These valves were able to stop the flow at room temperature, and allow the flow by opening within 20 s after heating was initiated using a Peltier element located just under the valve. The valve was successfully closed, thereby stopping the flow, and opened by heating. Although a detailed evaluation of the fluid behavior is necessary, we have demonstrated that our thermoresponsive valve can be opened and closed reversibly by temperature control. We believe that this thermoresponsive valve could potentially be used to control the flow of multiple reagents in µPADs.