Table 1.
Antibodies and blocking reagents for Western blot detection of aggregated human α-synuclein, amyloid-β and tau
| 1 st Antibody | Provider and product code | Epitope | Dilution | Blocking solution | 2 nd Antibody | Dilution |
|---|---|---|---|---|---|---|
| LB 509 (monoclonal) | Abcam, Cambridge, UK ab27766 | aa 115-122 of α-synuclein | 1:5,000 | 0.03% (w/v) Casein in TBS with 0.05% (w/v) Tween 20 | Alkaline-phosphatase-conjugated goat anti-mouse IgG (DAKO) | 1:5,000 |
| Phospho-S129 (monoclonal) | Abcam, Cambridge, UK ab51253 | α-synuclein phosphorylated at Ser129 | 1:2,000 | 3% (w/v) Low-fat milk powder in TBS with 0.05% (w/v) Tween 20 | Alkaline-phosphatase-conjugated goat anti-rabbit IgG (DAKO) | 1:10,000 |
| Anti-tau (monoclonal) | Merck Millipore, Darmstadt, Germany MAB2239 | C-terminal region of tau | 1:500 | 3% (w/v) Low-fat milk powder in TBS with 0.05% (w/v) Tween 20 | Alkaline-phosphatase-conjugated goat anti-mouse IgG (DAKO) | 1:10,000 |
| AT8 (monoclonal) | Thermo Scientific, Rockford, USA MN1020 | Phospho-PHF-tau (Ser202/Thr205) | 1:500 | 0.03% (w/v) Casein in TBS with 0.05% (w/v) Tween 20 | Alkaline-phosphatase-conjugated goat anti-mouse IgG (DAKO) | 1:10,000 |
| 6E10 (monoclonal) | Covance, Emeryville, USA SIG-39320 | aa 3-8 of Aβ | 1:2,000 | 5% (w/v) Low-fat choco powder in TBS with 0.05% (w/v) Tween 20a | Alkaline-phosphatase-conjugated goat anti-mouse IgG (DAKO) | 1:10,000 |
| 4G8 (monoclonal) | Covance, Emeryville, USA SIG-39220 | aa 17-24 of Aβ | 1:1,000 | 5% (w/v) Low-fat choco powder in TBS with 0.05% (w/v) Tween 20a | Alkaline-phosphatase-conjugated goat anti-mouse IgG (DAKO) | 1:10,000 |
aTina Zimmermann, personal communication and diploma thesis (2012, Establishment of the organotypic slice culture assay as a model for neurodegenerative diseases. Free University of Berlin, Berlin, Germany).