Table 6.
The bioanalytical techniques employed to determine α-synuclein oligomerization, fibrillation, and cytotoxicity.
| Biochemical/biophysical techniques | Events monitored in the system | References |
|---|---|---|
| Fluorescence polarization technique | α-syn aggregation | Luk et al., 2007 |
| Scanning for intensely fluorescent targets and atomic force microscopy | α-syn oligomers | Kostka et al., 2008 |
| High performance liquid chromatography (HPLC), Circular dichroism (CD), Fourier transform infrared spectroscopy (FTIR), Size exclusion HPLC, small-angle X-ray scattering, and atomic force microscopy (AFM) | HPLC (stability), Fourier transform infrared spectroscopy and atomic force microscopy (oligomer stabilization and fibrillation), CD (structural assessment) | Hong et al., 2008 |
| Extrinsic multiple-emission probe 4′-(diethylamino)-3-hydroxyflavone spectroscopy | Amyloid fibril formed by mutant α-syn | Celej et al., 2009 |
| Peptide mapping, Mass spectrometric and Ultra-high-field Nuclear Magnetic Resonance analysis | α-syn dimerization and inhibitor binding | Yamaguchi et al., 2010 |
| Fluorescence spectroscopy, Thioflavin T (Thio T) assay and Transmission electron microscopy (TEM) | α-syn fibrillation and preformed α-syn | Ono and Yamada, 2006 |
| Thio T assay, Light scattering measurement, size-exclusion HPLC, AFM | α-syn aggregation | Zhu et al., 2013 |
| Membrane potential-sensitive bis-oxonol fluorescent dye, DiBAC4(3) bio-sensing system | Cytotoxicity of C-terminal truncated α-syn 119 (α-syn119) | Kim et al., 2013 |
| Thio T assay, AFM, Nuclear magnetic resonance, Vesicle leakage assay | Fibril disassembling (Thio-T assay) | da Silva et al., 2013 |
| Lipid vesicle permeabilisation assay | Membrane damage by α-syn aggregates | Caruana et al., 2012 |
| Confocal single-molecule fluorescence spectroscopy | α-syn oligomer formation | Caruana et al., 2011 |
| Circular dichroism spectroscopy, Transmission electron microscopy, Atomic force microscopy, and Nuclear magnetic resonance analysis and Electrophysiological assays | α-syn oligomerization, NMR (binding to the N-terminal of α-syn) | Takahashi et al., 2015 |
| Electrochemical and localized surface plasmon resonance (LSPR), Cyclic and differential pulse voltammetry using redox probe [Fe(CN)6], Thio T assay, Surface plasmon resonance imaging, Transmission electron microscopy | α-syn oligomers by electrochemical and LSPR and Voltametry to detect binding of inhibitors to α-syn | Cheng et al., 2015 |
| Split firefly luciferase complementation assay with bioluminescence imaging | Visualizes oligomerization of α-syn in cell culture, striatum and SNc | Aelvoet et al., 2014 |
| ThT assays, Circular dichroism, Turbidity, and Rayleigh scattering measurements, Atomic force microscopy and Transmission electron microscopy | α-syn fibril formation | Fazili and Naeem, 2015 |
| CCK-8 staining on MPP(+)-induced SH-SYSY cells and Transmission electron microscopy, AO staining and western blotting in cells | Survival rate (CCK-8), autophagy (TEM), AO staining (lysosome), western (α-syn) | Wang et al., 2014 |