Guy-Crotte O, Barthe C, Figarella C
Groupe de Recherche sur les Glandes Exocrines, Marseille, France.
Electrophoresis. 1989 Aug-Sep;10(8-9):628-32. doi: 10.1002/elps.1150100816.
For the detection of the cystic fibrosis protein (CFP) in serum of cystic fibrosis (CF) carriers, thin-layer polyacrylamide gel isoelectric focusing proved inappropriate as a diagnostic test, but was useful for screening fractions on purification of CFP by chromatofocusing on a Mono P column. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis an Mr 12,000 protein (P12) was found in most CFP-positive sera, indicating good correlation between these two CF-associated proteins. Detection of the P12 protein by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was well reproducible and less delicate than IEF. The technique was also used to purify P12 from serum by two successive preparative electrophoresis steps in a 7.5-15% gradient and 15% homogeneous gel. The use of silver staining revealed that P12, which was present in all sera of CF patients and carriers with variable intensities, was also present in trace amounts in normal sera.
为检测囊性纤维化（CF）携带者血清中的囊性纤维化蛋白（CFP），薄层聚丙烯酰胺凝胶等电聚焦被证明不适用于诊断测试，但在通过Mono P柱进行层析聚焦纯化CFP时，对筛选组分很有用。在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳中，在大多数CFP阳性血清中发现了一种分子量为12,000的蛋白质（P12），表明这两种与CF相关的蛋白质之间具有良好的相关性。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳检测P12蛋白具有良好的重复性，且比等电聚焦操作更简便。该技术还用于通过在7.5 - 15%梯度凝胶和15%均一凝胶中进行两步连续制备电泳从血清中纯化P12。银染结果显示，P12存在于所有CF患者和携带者的血清中，强度各异，在正常血清中也有微量存在。
