Petruşcă D N, Popa O M, Gavrilă L, Sulică A
St. S. Nicolau Institute of Virology, Center of Immunology, Bucharest.
Rom J Virol. 1999 Jan-Dec;50(1-4):33-41.
Given the possibility that cell kinetics and p53 status may be important determinants of chemotherapeutical efficacy, the aim of the present study was to determine the optimal methods and conditions for qualitative and quantitative intracellular proteins detection. Bradford assay is the better choice for protein concentration detection because it is more sensitive and more rapid than Sheffield assay despite the fact that it utilizes a higher amount of samples. The direct staining method for flow-cytometrical detection of intracellular proteins is more rapid as compared to the indirect staining one, also providing information about protein expression during cell cycle phases, but it is low sensitive for protein expression estimation and is prohibitive for masked intracellular proteins like PCNA. More than that, it can be performed with both fixed and freshly isolated cells as compared to the indirect staining method, but the last one provides advantages by signal amplification and by its availability of using it for a large number of intracellular proteins detection.
鉴于细胞动力学和p53状态可能是化疗疗效的重要决定因素，本研究的目的是确定定性和定量检测细胞内蛋白质的最佳方法和条件。 Bradford法是蛋白质浓度检测的更好选择，因为尽管它使用的样品量较多，但比Sheffield法更灵敏、更快速。与间接染色法相比，流式细胞术检测细胞内蛋白质的直接染色法更快，还能提供细胞周期各阶段蛋白质表达的信息，但它对蛋白质表达估计的灵敏度较低，对于像增殖细胞核抗原（PCNA）这样的隐蔽细胞内蛋白质检测受限。此外，与间接染色法相比，它既可以用固定细胞也可以用新鲜分离的细胞进行检测，但间接染色法通过信号放大以及可用于大量细胞内蛋白质检测而具有优势。
