Simonin F, Briand J P, Muller S, de Murcia G
Institut de Biologie Moléculaire et Cellulaire du CNRS, Université Louis Pasteur, Strasbourg, France.
Anal Biochem. 1991 Jun;195(2):226-31. doi: 10.1016/0003-2697(91)90321-j.
We have recently devised an activity-blot procedure permitting the detection, on the same nitrocellulose sheet, of the functional poly(ADP ribose) polymerase (PARP) activity as well as the immunostained active peptide(s) after renaturation of the transferred protein(s). Using this technique we have analyzed the PARP activity in higher and lower eukaryotes directly on crude extracts from cell cultures. This procedure has been extended also to in situ screening of bacterial colonies expressing the PARP enzymatic activity.
我们最近设计了一种活性印迹法,可在同一张硝酸纤维素膜上检测功能性聚(ADP核糖)聚合酶(PARP)的活性,以及转移蛋白复性后免疫染色的活性肽。利用这项技术,我们直接对细胞培养物的粗提物分析了高等和低等真核生物中的PARP活性。该方法还扩展到了对表达PARP酶活性的细菌菌落进行原位筛选。