Liang Heng,Xing Jianyu,Liu Xicheng,Huang Liming
(School of Life Science and Technology,Xi'an Jiaotong University,Xi'an 710049,China)
Abstract:After investigating of the sample storing£¬lysis solution,IPG gel rehydration and staining etc. in two-dimensional gel electrophoresis(2-DE) experiments of the rat kidney tissue£¬it is found that even stored in -73 ¡æ£¬the same old samples are needed for comparing their 2-DE maps£®By many 2-DE experiments we obtained suitable lysis solution £¨Urea 9.8 mol/L£¬Tris 40 mmol/L£¬CHAPS 40 g/L£¬DTT 23 mmol/L£¬CA 5 g/L£¬PMSF 1 mmol/L and EDTA 1 mmol/L£© for the rat kidney tissue. The active inª²gel rehydration is helpful to the separation of large molecular weight proteins in the IPG gel£¬but increases the danger of small molecular weight protein running out from it£® During the process of staining£¬sufficiently washing steps ensures the lighter background and clear protein spots£® As a result£¬we achieved preferable separation in the high molecule district and alkaline section£¬and got a more satisfactory 2-DE pattern of the rat kidey tissue with 1 491 spots than that obtained in the references£®
Keywords:rat kidney-tissue;two-dimensional electrophoresis;proteome