Methods

• Protein biochemistry: recombinant protein production in prokaryotic (E. coli) and eukaryotic (D. discoideum, Baculovirus/Sf9 insect cells) systems; isolation and purification of native proteins from various tissues (skeletal and heart muscle, brain, other tissues); preparative ultracentrifugation; chromatography (affinity chromatography, ion exchange chromatography, size exclusion chromatography, hydrophobic interaction chromatography) on AEKTApure and BioRad NGC column chromatography systems
• Molecular biology: molecular cloning; PCR; plasmid generation
• Protein Characterization: plate-reader assisted steady-state enzyme kinetics; fast transient kinetics (stopped-flow, quench flow); quantification of protein-protein interactions (Microscale thermophoresis, Fluorescence polarization/ anisotropy; Circular Dichroism, Calorimetry – ITC and DSC)
• Fluorescence microscopy based functional assays: TIRF; in vitro motility assays; actin-polymerization, depolymerization and bundling assays
• Cell Biology: mammalian cell culture; transfection; stable and transient gene expression; gene silencing (siRNA- or shRNA-mediated knock-down); CRISPR/Cas-mediated gene editing; cell toxicity assays
• Microscopy: epifluorescence and TIRF microscopy of fluorescently labelled proteins, protein complexes and cells; Confocal Microscopy; FRAP; FRET; Live Cell Imaging; negative stain electron microscopy