Bekei Beata, Rose Honor May, Herzig Michaela, Stephanowitz Heike, Krause Eberhard, Selenko Philipp
Department of NMR-assisted Structural Biology, Leibniz Institute of Molecular Pharmacology (FMP Berlin), Berlin, Germany.
Methods Mol Biol. 2012;895:67-83. doi: 10.1007/978-1-61779-927-3_6.
Irrespective of how isotope-labeled proteins are delivered into mammalian cells, laboratory routines are needed to assess the quality of the resulting in-cell NMR samples. These include methods to evaluate overall cell viability, protein transduction efficiency, intracellular protein concentration, localization, and stability. In addition, quality control experiments to assess protein leakage from manipulated cells are of particular importance for in-cell NMR experiments. The purpose of this chapter is to outline qualitative and quantitative methods to determine general biological properties of in-cell NMR samples in order to ensure the highest possible standards for in-cell NMR studies.
无论同位素标记的蛋白质是如何导入哺乳动物细胞的，都需要实验室常规方法来评估所得细胞内核磁共振（NMR）样品的质量。这些方法包括评估细胞总体活力、蛋白质转导效率、细胞内蛋白质浓度、定位和稳定性的方法。此外，评估经处理细胞中蛋白质泄漏情况的质量控制实验对于细胞内NMR实验尤为重要。本章的目的是概述定性和定量方法，以确定细胞内NMR样品的一般生物学特性，从而确保细胞内NMR研究达到尽可能高的标准。
