The invention relates to a method of detecting the content of N1,N<12>-diacetylspermine (called as DAS for short), and particularly relates to a kit used for detecting the content of the N1,N<12>-diacetylspermine (DAS), preparing methods of an antigen used in the kit and an antibody used in the kit, a developing method of the kit, and a preparing scheme of the kit. The kit optimizes detecting methods of the content of the DAS. Compared with the prior art, the kit is advantaged by simple and convenient operation, a high detection speed, high precision, good repeatability, capability of allowing detection results to be not influenced by operation, capability of being used for large-scale sample testing, and the like.