两种不同溶液体系提取大鼠脊髓蛋白双向电泳图谱比较

目的：比较尿素／CHAPS和尿素／硫脲／CHAPS／SB3－10两种不同溶液体系提取大鼠脊髓蛋白双电泳（two-dimensional electrophoresis,2-DE)图谱的差异。方法：大鼠脊髓经三氯醋酸／丙酮沉淀粉蛋白后，分别用尿素／CHAPS和尿素／硫脲／CHAPS／SB3－10两种不同的溶液体系提取蛋白。以固相PH梯度（IPG）等电聚焦为第一向，SDS－PAGE水平电泳为第二向进行蛋白质2－DE。图像分析软件ImageMaster 2D Elite3.01分析蛋白质2－DE图谱。结果：两种蛋白提取溶液分别获得1059和1023个蛋白（亚基）斑点，其中790个蛋白在两张图谱中重叠，其余269和233个蛋白点为两种体系各自所特有。两种提取方法共获得1292个蛋白斑点。结论：综合使用不同蛋白提取方法有助于提高组织蛋白2－DE图谱的完整性。

Comparison of the two dimensional electrophoresis maps of rat spinal cord protein extracted by two different solution systems

Objective:To compare the two dimensional electrophoresis(2 DE) maps of rat spinal cord protein extracted by two different solution systems.Methods: Adult rat spinal cord protein was precipitated with 10% trichloracetic acid in acetone and resuspended in 8 mol/L urea plus 4%CHAPS (A solution) or, 5 mol/L urea, 2 mol/L thiourea, 2%CHAPS plus 2%SB3 10 (B solution). One hundred and fifty micrograms of protein was loaded on 18 cm IPG strip holder and run isoelectric focusing electrophoresis as the first dimension, then horizontal SDS PAGE as the second dimension. Protein spots were visualized by silver stain.Results:There were 1 059 and 1 023 protein spots in each map, of which 790 spots were matched in two maps. There were 269 and 233 spots exclusively extracted by A and B solutions, respectively. Taken together, 1292 different spots were totally obtained by A and B solutions.Conclusion: Integrating protein spots extracted by different solution systems is beneficial for achieving intact 2 DE map of tissues.