CAJ | 학술논문

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向电泳（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 Elite 3.01分析蛋白质2-DE图谱。结果：两种蛋白提取溶液分别获得1 059和1 023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1 292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。
목적:비교뇨소/CHAPS화뇨소/류뇨/CHAPS/SB3-10량충불동용액체계제취대서척수단백쌍향전영（two-dimensional electrophoresis, 2-DE）도보적차이。방법：대서척수경삼록작산/병동침정단백후，분별용뇨소/CHAPS화뇨소/류뇨/CHAPS/SB3-10량충불동적용액체계제취단백。이고상pH제도（IPG）등전취초위제일향，SDS-PAGE수평전영위제이향진행단백질2-DE。도상분석연건ImageMaster 2D Elite 3.01분석단백질2-DE도보。결과：량충단백제취용액분별획득1 059화1 023개단백（아기）반점，기중790개단백재량장도보중중첩，기여269화233개단백점위량충체계각자소특유。량충제취방법공획득1 292개단백반점。결론：종합사용불동단백제취방법유조우제고조직단백2-DE도보적완정성。
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.