Heat transfer performances of a pulsating heat pipe (PHP) having internal and external diameter with 4.5 mm and 6 mm with various contents of refrigerant are experimentally investigated. The working fluid as R404A refrigerant was filled in the volume ratios from 0% to 80% and the heat input was controlled in the range from 10 W to 80 W. Obtained results exhibited the ability of R404A refrigerant can enhance the thermal performance in steady state condition. The average temperature difference of the evaporating section and condensing section in the 80% filling volume ratio decreased from 9.5 °C to 2.5 °C when the heating power increase from 10 W to 80 W. The thermal resistance of evaporator and condenser decreased with an increase of the heat input as well. For other filling volume ratios, the trend of temperature difference and thermal resistance was similar to that of the 80% volume filling ratio. Considering the same heat input, the highest heat transfer performance was found at the 80% volume filling ratio. Refrigerant with a relatively low dynamic consistency can lead to relatively high velocity in the PHP that can reduce the temperature difference between the evaporating section and condensing section.