Zakharian Research Methods

Research Methods

Protein Biochemistry

  • Protein isolation and purification: from mammalian or bacterial cell cultures
  • Sub-cellular protein fractionation, immunoblotting and immunoprecipitation
  • Mass spectral analysis
  • Analysis of posttranslational modifications
  • Biotinylation assays

Electrophysiology

  • Planar Lipid Bilayer measurements
  • Whole-cell patch clamp recordings on mammalian cells

Fluorescence

  • Calcium imaging, dual excitation with fura-2
  • Confocal microscopy
  • Immunocytochemistry and Immunohistochemistry

Molecular Biology

  • Transfection of mammalian cells
  • Mutation of cDNA to study recombinant ion channels
  • Real-time PCR
  • Chromatin immunoprecipitation (ChIP)

Cell Cultures

  • Cultured and primary mammalian cells, those include: HEK-293 cells, prostate cancer cells LnCAP, DU145 and PC3, prostate epithelial cells RWPE1 and RWPE2, dorsal root ganglion (DRG) neurons, hippocampal neurons, glioma cells U87 and U251, and lung cancer cells H1299
  • Establishment of stable cells expressing TRPM8 (Myc, His-tagged), polyphosphatase scPPX
  • Cell cycle analysis by fluorescence-activated cell sorting (FACS)