The detection of human serum albumin (HSA) in bodily fluids is of great significance in the biomedical area because HSA in bodily fluids is commonly used as a biomarker for the early diagnosis of diseases. To detect HSA, we employed HDBB, 4,4'-(hydrazine-1,2-diylidene bis(methanylylidene)) bis(3-hydroxybenzoic acid), as a fluorescent probe with a large Stokes shift. HDBB had obvious excited state intramolecular proton transfer (ESIPT) and aggregation-induced emission (AIE) features. We elucidated the ESIPT characteristics of HDBB through the DFT approach. We also performed a molecular docking simulation between HDBB and HSA, showing that HDBB primarily bonded to HSA via hydrophobic force and hydrogen bonds. The FL intensities of HDBB with HSA concentrations had a linear range of 0.01-0.2 mg mL-1 (R2 = 0.9995), and the LOD was 1.104 μg mL-1. We also used the probe to detect HSA in urine, with spiked recoveries of 98.10-105.33%. Given its high selectivity and feasible synthesis, HDBB has potential applications in detecting HSA in real biological systems.