To comprehensively and quickly identify proteins of mouse liver, the tryptic peptides were pre-frac-tioned and enriched by isoelectric focusing technigue using Agilent 3100 OFFGEL Fractionator into 24 frac-tions, followed by RPLC and MS/MS analysis. Each MS/MS spectrum was searched for mouse species against the Swissprot database. The results were analyzed which resulted in the identification of proteins in mouse liver. Finally, 3474 proteins were identified from mouse liver, 832 of them were new identified proteins. Bioinformatic analyses were performed to demonstrate the physicochemical properties, cellular locations and functions of the proteins. The combined use of liquid fractionation and LC-MS allows a considerable increase in proteome coverage of very complex samples prepared from the tissues extracts and supports the low-concen-trated protein identification. This study can provide useful data to research C57 mouse liver proteins.%利用液相等电聚焦预分离技术结合液相色谱-质谱( LTQ-Orbitrap)联用技术，研究了C57小鼠肝脏的蛋白质表达谱.质谱分析结果采用MaxQuant1.4.1.2软件搜索数据库，共鉴定出3474个蛋白(2个以上唯一肽段).用DAVID在线工具、GO分类工具和IPA软件对鉴定蛋白进行生物信息学分析，单独发现832个新蛋白.研究结果表明，通过基于等电点和亲疏水性的两维分离，提高了质谱鉴定蛋白数，可以鉴定出更多低丰度蛋白.
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