OBJECTIVE: To establish a urinary proteomic map on two-dimensional polyacrylamide gelelectrophoresis (2-DE) in children with idiopathic nephrotic syndrome (INS). METHODS: The proteins from INS children were purified by four various means, separated by 2-DE, and stained by silver. RESULTS The sequential preparation of urinary proteins by acetone precipitation and dislysis, when the sample was 300 μg, resulted in a clear background, well-resolved and reproducible 2-DE urinary protemic map in children with INS. CONCLUSIONS A steady 2-DE technique for urinary protemic map in children with INS was established, which can be effectively applied in urinary proteomics of the disease.［Chin J Contemp Pediatr, 2007, 9 (2):122-124］