In-cell NMR in mammalian cells: part 3

Methods Mol Biol. 2012:895:67-83. doi: 10.1007/978-1-61779-927-3_6.

Abstract

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.

MeSH terms

  • Amino Acid Sequence
  • Blotting, Western
  • Flow Cytometry
  • HeLa Cells
  • Humans
  • Mass Spectrometry
  • Microscopy, Fluorescence
  • Molecular Sequence Data
  • Nuclear Magnetic Resonance, Biomolecular*
  • Protein Conformation
  • Staining and Labeling
  • Tandem Mass Spectrometry
  • Trypan Blue / chemistry
  • alpha-Synuclein / chemistry*

Substances

  • alpha-Synuclein
  • Trypan Blue