Proteome research of Human renal tubular epithelial cell line HK-2 by two-dimensional gel electrophoresis and computer image analysis

Objective Two-dimensional gel electrophoresis(2DGE)is a key technique for proteomics. The aim of the research is to establish the two-dimensional gel electrophoresis and computer assisted image analysis for the proleome research of human renal tubular epithelial cell. Methods The human renal tubular epithelial cell line HK-2 was used as model. The rehydrated IPG strips containing protein samples were isoelectric focusing(IEF) . After IEF,the equilibrated strips were transferred to the vertical SDS gel. The silver stained gels were scanned by the densitometer,which generated the digitalized images. Using new generation two-dimensional image analysis software,ImageMaster 2D Elite 3.01 ,the 2DGE of proteins extracted from cultured renal tubular epithelial cell was processed. The analysis procedure,including image acquirement,automatic spot detection and quantification,analysis results report,was discussed. The method and volume of loading sample,choice of immobilized pH gradient(IPG)gels,concentration of SDS gels,preset of electric parameters,protocol for staining were improved. Results Using the proper method described above,satisfactory 2DGE maps of HK-2 was obtained and analysis results were reported. Conclusions We have used proteomic tools to construct protein maps of human renal tubular epithelial cell,the technique will be used in renal research to characterize physiological processes and disease.