采用非涂层毛细管,以150mm o l/L硼酸盐缓冲液为电泳缓冲液,30g/L聚乙二醇(PEG)20000为筛分介质,经对分离条件进行优化,成功地建立了用无胶筛分毛细管电泳检测微量蛋白质的方法。用所建立的方法测定骨桥蛋白,其批内、批间迁移时间的相...采用非涂层毛细管,以150mm o l/L硼酸盐缓冲液为电泳缓冲液,30g/L聚乙二醇(PEG)20000为筛分介质,经对分离条件进行优化,成功地建立了用无胶筛分毛细管电泳检测微量蛋白质的方法。用所建立的方法测定骨桥蛋白,其批内、批间迁移时间的相对标准偏差均小于5%,回收率大于95%,被检测样品中骨桥蛋白的含量与其峰面积呈良好的线性关系(相关系数为0.996),最低检测限为0.079g/L。考察了无血清饥饿培养对血管平滑肌细胞分泌骨桥蛋白的影响,结果表明无血清饥饿培养24h骨桥蛋白的合成与分泌达到高峰,此后随时间延长含量随之降低,此结果与采用W estern b lo t方法检测的结果一致。该方法具有进样量小(nL级)、检测速度快、可自动化等优点,是一种简便、快捷的检测微量蛋白质的好方法。展开更多
在分离Y染色体性别决定区(Sex determining region of the Y chromosome,SRY)、人11号染色体短臂β-珠蛋白基因部分序列(108/B)扩增产物和PBR322/HaeⅢ标准片段的混合样品时,SRY和108/B的迁移时间均产生较大的偏差,在尿素变性剂存在的...在分离Y染色体性别决定区(Sex determining region of the Y chromosome,SRY)、人11号染色体短臂β-珠蛋白基因部分序列(108/B)扩增产物和PBR322/HaeⅢ标准片段的混合样品时,SRY和108/B的迁移时间均产生较大的偏差,在尿素变性剂存在的无胶筛分体系中迁移时,这种迁移时间的偏差得以消除,这意味着尿素变性剂的存在消除了DNA空间结构对迁移的影响,DNA在此筛分介质中的迁移只与其片段长度有关.在变性无胶筛分体系中鉴别PCR产物,其结果的准确度明显提高.展开更多
Quasi-interpenetrating network of polyacrylamide （PAA） and polyvinylpyrrolidone （PVP） had been successfully used for single-base resolution of double-stranded DNA （0.76 for 123 bp/124 bp） and single-stranded DNA...Quasi-interpenetrating network of polyacrylamide （PAA） and polyvinylpyrrolidone （PVP） had been successfully used for single-base resolution of double-stranded DNA （0.76 for 123 bp/124 bp） and single-stranded DNA fragments （0.97 for 123 b/124 b） with UV detection. This quasi-IPN （interpenetrating network） sieving matrix showed low viscosity （23.5 mPa·s at 25 ℃） and decreased with increasing temperature. This polymer also exhibited dynamically coating capacity and could be used in the uncoated capillary. The effects of temperature and electric field strength on the DNA separation of quasi-IPN matrix were also investigated and found that the temperature and electric field strength could markedly affected the mobility behavior of DNA fragments. This polymer matrix has also applied to separate the bigger DNA fragments by capillary electrophoresis with UV detection. Under the denaturing conditions, this matrix separated the samples with last fragment of 1353 base in 40 rain, in which the doublet of 309/310 base was partial separated and the resolution was 0.88.展开更多
文摘采用非涂层毛细管,以150mm o l/L硼酸盐缓冲液为电泳缓冲液,30g/L聚乙二醇(PEG)20000为筛分介质,经对分离条件进行优化,成功地建立了用无胶筛分毛细管电泳检测微量蛋白质的方法。用所建立的方法测定骨桥蛋白,其批内、批间迁移时间的相对标准偏差均小于5%,回收率大于95%,被检测样品中骨桥蛋白的含量与其峰面积呈良好的线性关系(相关系数为0.996),最低检测限为0.079g/L。考察了无血清饥饿培养对血管平滑肌细胞分泌骨桥蛋白的影响,结果表明无血清饥饿培养24h骨桥蛋白的合成与分泌达到高峰,此后随时间延长含量随之降低,此结果与采用W estern b lo t方法检测的结果一致。该方法具有进样量小(nL级)、检测速度快、可自动化等优点,是一种简便、快捷的检测微量蛋白质的好方法。
文摘在分离Y染色体性别决定区(Sex determining region of the Y chromosome,SRY)、人11号染色体短臂β-珠蛋白基因部分序列(108/B)扩增产物和PBR322/HaeⅢ标准片段的混合样品时,SRY和108/B的迁移时间均产生较大的偏差,在尿素变性剂存在的无胶筛分体系中迁移时,这种迁移时间的偏差得以消除,这意味着尿素变性剂的存在消除了DNA空间结构对迁移的影响,DNA在此筛分介质中的迁移只与其片段长度有关.在变性无胶筛分体系中鉴别PCR产物,其结果的准确度明显提高.
文摘Quasi-interpenetrating network of polyacrylamide （PAA） and polyvinylpyrrolidone （PVP） had been successfully used for single-base resolution of double-stranded DNA （0.76 for 123 bp/124 bp） and single-stranded DNA fragments （0.97 for 123 b/124 b） with UV detection. This quasi-IPN （interpenetrating network） sieving matrix showed low viscosity （23.5 mPa·s at 25 ℃） and decreased with increasing temperature. This polymer also exhibited dynamically coating capacity and could be used in the uncoated capillary. The effects of temperature and electric field strength on the DNA separation of quasi-IPN matrix were also investigated and found that the temperature and electric field strength could markedly affected the mobility behavior of DNA fragments. This polymer matrix has also applied to separate the bigger DNA fragments by capillary electrophoresis with UV detection. Under the denaturing conditions, this matrix separated the samples with last fragment of 1353 base in 40 rain, in which the doublet of 309/310 base was partial separated and the resolution was 0.88.
