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毛细管电泳检测肺癌及癌旁正常组织蛋白质混合物差异
  • ISSN号:0567-7351
  • 期刊名称:《化学学报》
  • 时间:0
  • 分类:Q503[生物学—生物化学]
  • 作者机构:[1]兰州军区兰州总医院临床药理基地.兰州730050, [2]兰州大学生命科学学院,兰州730000, [3]兰州大学药学院,兰州730000, [4]西北师范大学生命科学学院,兰州730070
  • 相关基金:国家自然科学基金(Nos20775089,30970312)资助项目.
中文摘要:

以建立的毛细管电泳(cE)-激光诱导荧光(LIF)检测蛋白质的方法对提取肺癌及癌旁正常组织蛋白质混合物(变性/活性)差异进行检测.采用异硫氰酸荧光素(FITC)为衍生剂,电泳缓冲液为l×TBE(TBE为Tris-硼酸-EDTA,变性电泳pH10.0,活性电泳为pH8.3且含有2mg/L考马斯亮蓝),分离电压15kV,柱温15℃,电动进样(10kV×10s),激发波长,发射波长=488/520nm检测时,肺癌及癌旁正常组织蛋白质混合物样品得到较好分离且有明显差异.与目前常用蛋白分析方法:变性SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)以及活性蓝绿温和胶电泳(BN-PAGE)进行比较.BN—PAGE结果显示肺癌组织相比正常组织有较明显蛋白种类差异;SDS—PAGE结果表明一些蛋白质表达量差异也是肺癌及癌旁正常组织的显著差别,且主要集中在20~l16kDa,CE—LIF检测结果与PAGE结果大致相同,且CE—LIF检测蛋白质的灵敏度高于PAGE,能更准确反映肺癌及癌旁正常组织的蛋白质差异.结论是CE—LIF可用于蛋白质差异检测,时间短,效果较好,对活性蛋白质进行分析体现了其优点:可提供较强的动力——电压,及强动力下良好的温度稳定性.

英文摘要:

Protein mixture (denatured and native) differences of extracted from lung cancer and adjacent normal tissue by established method of capillary electrophoresis (CE)-laser-induced fluorescence (LIF) detection of protein was detected using the fluorescein isothiocyanate (FITC) as a derivative agent. When detected by the 1 × TBE electrophoresis buffer solution (pH 10.0 of denaturing gel electrophoresis and pH 8.3 of native gel electrophoresis containing 2 mg/L Coomassie brilliant blue), separation voltage of 15 kV, column temperature of 15 ℃, electric injection (10 kV X 10 s), and excitation wavelength/emission wavelength of 488/520 nm, lung cancer and adjacent normal tissue protein mixture samples obtained a better separation and there was significant difference. Comparing with the current commonly used analysis methods, the SDS-denatured polyacrylamide gel electrophoresis (SDS-PAGE) as well as the Blue Native polyacrylamidegel electrophoresis (BN-PAGE), the BN-PAGE results showed that compared to normal tissue, the lung cancer tissue had more significant protein type differences. But the SDS-PAGE results showed that some differences of protein expression had significant difference between the lung cancer and adjacent normal tissue, and mainly concentrated in 20~116 kDa. CE-LIF detection results were roughly the same with PAGE, and CE-LIF is more sensitive than PAGE and it can more accurately reflect protein differences of lung cancer and adjacent normal tissue. We concluded that the CE-LIF could be used to detect protein differences. And the time was shorter, effect was better, and analysis of active protein reflected its advantage of providing a strong driving force-voltage and a well temperature stability by strong motivation.

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期刊信息
  • 《化学学报》
  • 北大核心期刊(2014版)
  • 主管单位:中国科学院
  • 主办单位:中国化学会 中国科学院上海有机化学研究所
  • 主编:周其林
  • 地址:上海市零陵路345号
  • 邮编:200032
  • 邮箱:hxxb@sioc.ac.cn
  • 电话:021-54925085
  • 国际标准刊号:ISSN:0567-7351
  • 国内统一刊号:ISSN:31-1320/O6
  • 邮发代号:4-209
  • 获奖情况:
  • 首届国家期刊奖,第二届国家期刊奖提名奖,中国期刊方阵“双高期刊”
  • 国内外数据库收录:
  • 俄罗斯文摘杂志,美国化学文摘(网络版),荷兰文摘与引文数据库,美国科学引文索引(扩展库),日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),英国英国皇家化学学会文摘,中国北大核心期刊(2000版)
  • 被引量:28694