Hinson W G, Pipkin J L, Hudson J L, Anson J F, Tyrer H
Cytometry. 1982 May;2(6):390-4. doi: 10.1002/cyto.990020606.
Biochemical analysis of proteins extracted from mammalian cell nuclei which were sorted according to DNA content was greatly simplified using a special collection device to recover sorted nuclei. The device was developed for use with a sorting flow cytometer to provide sufficient numbers of particles to achieve adequate sensitivity and resolution while minimizing sample degradation and loss. The design, evaluation and application of the collection device were described with emphasis on sample preparation. Nuclear populations G0 + G1, S, and G2 + M, were sorted form regenerating rat liver tissue preparations and collected separately in the special chambers. Nuclear proteins were extracted and analyzed by polyacrylamide gel isoelectric focusing or capillary isotachophoresis.
使用一种特殊的收集装置来回收分选的细胞核，极大地简化了对根据DNA含量分选的哺乳动物细胞核中提取的蛋白质进行的生化分析。该装置是为与分选流式细胞仪配合使用而开发的，以提供足够数量的颗粒，在尽量减少样品降解和损失的同时实现足够的灵敏度和分辨率。重点介绍了样品制备，描述了收集装置的设计、评估和应用。从再生大鼠肝脏组织制备物中分选核群体G0 + G1、S和G2 + M，并分别收集在特殊腔室中。通过聚丙烯酰胺凝胶等电聚焦或毛细管等速电泳提取和分析核蛋白。
