The invention provides a thermo-setting transfer mould release agent. The thermo-setting transfer mould release agent comprises polyisocyanates and an ingredient, wherein the ingredient comprises the following components in part by weight: 15 to 45 parts of fluorocarbon resin, 2 to10 parts of polytetrafluoroethylene micro powder, 40 to 62 parts of C4-C8 alkane, 0.2 to 0.5 part of antifoaming agent, 1 to 3 parts of flatting agent, 1 to 5 parts of release auxiliaries, and 1 to 4 parts of flatting silica, wherein the weight ratio of the polyisocyanates to the fluorocarbon resin is 1 to 8. and the method for preparing and using the mould release agent is as follows: adding the fluorocarbon resin while agitating the C4-C8 alkane and the polytetrafluoroethylene; adding the antifoaming agent, the flatting agent, the release auxiliaries and the flatting silica, and then agitating the mixture; and, regulating the weight ratio of the fluorocarbon resin to the polyisocyanates in the product to be 8 to 1. The acquired mould release agent is coated on a PET (Polymer) film by a mesh plate and then dried for the use. The mould release agent is not transferred after being transfer-printed, so that the performances of the printing oil are not affected; the bottom film has no change after transfer-printing, and the friction resistant and solvent resistant performances are very good.