A die seal ring and a manufacturing method thereof are provided. The die seal ring includes a substrate, a dielectric layer, and conductive layers. The dielectric layer is disposed on the substrate. The conductive layers are stacked on the substrate and located in the dielectric layer. Each of the conductive layers includes a first conductive portion and a second conductive portion. The second conductive portion is disposed on the first conductive portion. A width of the first conductive portion is smaller than a width of the second conductive portion. A first air gap is disposed between a sidewall of the first conductive portion and the dielectric layer. A second air gap is disposed between a sidewall of the second conductive portion and the dielectric layer. The die seal ring and the manufacturing method thereof can effectively prevent cracks generated during the die sawing process from damaging the circuit structure.